iGEM 2015 — Team NCTU_Formosa

# Team:NCTU Formosa/Notebook

* [Home](https://2015.igem.org/Team%3ANCTU_Formosa)
* Project
  + [Description](https://2015.igem.org/Team%3ANCTU_Formosa/Description)
  + [Design](https://2015.igem.org/Team%3ANCTU_Formosa/Design)
  + [Results](https://2015.igem.org/Team%3ANCTU_Formosa/Results)
  + [Modeling](https://2015.igem.org/Team%3ANCTU_Formosa/Modeling)
  + [Safety](https://2015.igem.org/Team%3ANCTU_Formosa/Safety)
* [Parts](https://2015.igem.org/Team%3ANCTU_Formosa/Parts)
  + [Basic Parts](https://2015.igem.org/Team%3ANCTU_Formosa/Basic_Part)
  + [Composite Parts](https://2015.igem.org/Team%3ANCTU_Formosa/Composite_Part)
* Notebook
  + [Lab Safety](https://2015.igem.org/Team%3ANCTU_Formosa/LabSafety)
  + [Lab note](https://2015.igem.org/Team%3ANCTU_Formosa/Notebook)
  + [Protocol](https://2015.igem.org/Team%3ANCTU_Formosa/Protocol)
* Policy & Practices
  + [Human Practice](https://2015.igem.org/Team%3ANCTU_Formosa/Practices)
  + [Collaborations](https://2015.igem.org/Team%3ANCTU_Formosa/Collaborations)
* [Achievement](https://2015.igem.org/Team%3ANCTU_Formosa/Achievement)
* [Team](https://2015.igem.org/Team%3ANCTU_Formosa/Attributions)

Lab note

* [2015 April](#tab2)
* [2015 May](#tab3)
* [2015 June](#tab4)
* [2015 July](#tab5)
* [2015 August](#tab6)

|  |  |  |  |  |  |  |
| --- | --- | --- | --- | --- | --- | --- |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
|  |  |  | **1** | **2** | **3** | **4** |
| **5** | **6** | **7** | **8** | **9** | **10** | **11** |
| **12** | **13** | **14** | **15**   **Transformation of** GBP[FadL-GBP]  Cultivation of  OmpA[OmpA+pSB1K3] Kr | **16**   Cultivation of  GBP[FadL-GBP]   **Plasmid purification of** OmpA[OmpA+pSB1K3] Kr   **Digestion of** OmpA[OmpA+pSB1K3] Kr (EcoRI and PstI) | **17**  **Plasmid purification of**  GBP[FadL-GBP]   **Digestion of**  GBP[FadL-GBP] (EcoRI and PstI)   **Electrophoresis of** digested product:  GBP[FadL-GBP] (EcoRI and PstI) | **18**   **Transformation of** ligated product:  GBP[FadL-GBP+pSB1C3] Cr |
| **19**   **Cultivation of**  ligated product: GBP[FadL-GBP] Cr  PCR product: Terminator[J61048] | **20** | **21**   **Cultivation of**  GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar  OmpA[OmpA+pSB1C3] Cr  GBP[FadL-GBP+pSB1C3] Cr | **22**   **Plasmid purification of**  GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar  OmpA[OmpA+pSB1C3] Cr  GBP[FadL-GBP+PSB1C3] Cr   **Digestion of**  GFP[J23101+B0034+E0040+J61048] Ar (EcoRI and PstI)  OmpA[OmpA] Cr (EcoRI and PstI)  GBP[FadL-GBP] Cr (EcoRI and PstI)   **Electrophoresis of** digested product:  GFP[J23101+B0034+E0040+J61048] Ar (EcoRI and PstI)  OmpA[OmpA] Cr (EcoRI and PstI)  GBP[FadL-GBP] Cr (EcoRI and PstI) | **23**   **Digestion of** OmpA[OmpA] Cr (EcoRI and SpeI)  GBP[FadL-GBP] Cr (EcoRI and SpeI)   **Electrophoresis of** digested product:  OmpA[OmpA] Cr (EcoRI and SpeI)  GBP[FadL-GBP] Cr (EcoRI and SpeI) | **24**   **Digestion of**  OmpA[OmpA] Cr (XbaI and PstI)  GBP[FadL-GBP] Cr (XbaI and PstI) | **25**   **Electrophoresis of** digested product:  OmpA[OmpA] Cr (XbaI and PstI)  GBP[FadL-GBP] Cr (XbaI and PstI)   **Digestion of**  RBS[B0034] Ar (XbaI and PstI)  backbone[pSB1A3] Ar (EcoRI and PstI)  Terminator[J61048] Ar (XbaI and PstI)   **Electrophoresis of** digested product:  RBS[B0034] Ar (XbaI and PstI)  backbone[pSB1A3] Ar (EcoRI and PstI)  Terminator[J61048] Ar (XbaI and PstI)   **Ligation of**  OmpA[J23101+B0034+OmpA+pSB1A3] Ar  GBP[GBP+B0034+pSB1K3] Kr  OmpA[OmpA+J61048+pSB1K3] Kr |
| **26**   **Ligation of**  RFP[E1010+B0034+pSB1K3] Kr   **Transformation** of ligated product:  OmpA[J23101+B0034+OmpA+pSB1A3] Ar  GBP[FadL-GBP+B0034+pSB1K3] Kr  OmpA[OmpA+J61048+pSB1K3] Kr   **Electrophoresis of** digested product:  RFP[E1010+J61048+pSB1K3] Kr  [J23101+B0034] | **27**   **Digestion of**  Terminator[J61048] Ar (XbaI and PstI)   **PCR of** ligated product:  OmpA[J23101+B0034+OmpA+pSB1A3] Ar  OmpA[OmpA+J61048+pSB1K3] Kr | **28**   **Electrophoresis of**  RFP[E1010+J61048+pSB1K3] Kr  OmpA[J23101+B0034+OmpA+pSB1A3] Ar **Transformation of** ligated product:  GBP[B0034+FadL-GBP+pSB1A3] Ar  OmpA[OmpA+J61048+pSB1K3] Kr **PCR of** ligated product  OmpA[J23101+B0034+OmpA] Ar | **29**   **Cultivation of**  RFP[E1010+J61048+pSB1K3] Kr **PCR of**  OmpA[J23101+B0034+OmpA] Ar  **Electrophoresis of**  OmpA[J23101+B0034+OmpA] Ar **Ligation of**  GBP[FadL-GBP+J61048+pSB1K3] Kr | **30**   **Plasmid purification of**  RFP[E1010+J61048+pSB1K3] Kr **Ligation of**  OmpA[J23101+B0034+OmpA] Ar **PCR of**  OmpA[J23101+B0034+OmpA] Ar **Transformation of**  mini: Terminator[J61048]  ligated product: OmpA[J23101+B0034+OmpA] Ar |  |  |

|  |  |  |  |  |  |  |
| --- | --- | --- | --- | --- | --- | --- |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
|  |  |  |  |  | **1**  **PCR of** Transformation product:  mini: Terminator[J61048]  ligated product: OmpA[J23101+B0034+OmpA] Ar  GBP[B0034+FadL-GBP+pSB1A3] Ar   **Electrophoresis of**  mini: Terminator[J61048]  ligated product: OmpA[J23101+B0034+OmpA] Ar | **2** |
| **3**  **Ligation of**  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar | **4**  **Ligation of**  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar   5/4 **Transformation off** ligated products:  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar   **Digestion of**   RFP[E1010+J61048] (Kr) (XbaI and PstI)   **PCR of** ligated products:  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar   **Electrophoresis of** PCR products:  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar | **5**  **Electrophoresis of**  digested products:  RFP[E1010+J61048] (Kr) (XbaI and PstI)  OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI)  GBP[FadL-GBP+J61048] Kr (EcoRI and PstI)   **PCR of**  OmpA[J23101+B0034+OmpA+pSB1A3] Ar   **Cultivation of** PCR product:  RFP[E1010+J61048+pSB1K3] Kr  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar | **6**  **Plasmid purification of**  RFP[E1010+J61048+pSB1K3] Kr  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar   **Digestion of**   GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)   **Cultivation of** PCR product:  OmpA[J23101+B0034+OmpA+pSB1A3] Ar   **PCR of** backbone:  [pSB1C3] Cr  [pSB1A3] Ar  [pSB1K3] Kr   **Electrophoresis of**  PCR products: OmpA[J23101+B0034+OmpA+pSB1A3] Ar  digested products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) | **7**  **Digestion of**   RFP[E1010+J61048+pSB1K3] Kr (EcoRI and PstI)  RFP[E1010+J61048+pSB1K3] Kr (XbaI and PstI)  [pSB1C3] Cr (EcoRI and PstI)  [pSB1A3] Ar (EcoRI and PstI)  [pSB1K3] Kr (EcoRI and PstI)  OmpA[J23101+B0034+OmpA+pSB1A3] Ar (EcoRI and PstI)   **Plasmid purification of**  OmpA[J23101+B0034+OmpA+pSB1A3] Ar  [pSB1K3] Kr   **Electrophoresis of** digested products:  RFP[E1010+J61048+pSB1K3] Kr (EcoRI and PstI)    RFP[E1010+J61048+pSB1K3] Kr (XbaI and PstI)  [pSB1C3] Cr (EcoRI and PstI)  [pSB1A3] Ar (EcoRI and PstI)  [pSB1K3] Kr (EcoRI and PstI)   **Ligation of**  RFP[B0034+E1010+J61048+pSB1C3] Cr | **8**  **Electrophoresis of** digested products:  OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI)   **Transformation off** ligated products:  RFP[B0034+E1010+J61048+pSB1C3] Cr   **PCR of**  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar   **Electrophoresis of** PCR products:  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar   **Transformation off**  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar  [B0030] Ar  [J23110] Ar | **9**  **PCR of**  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar  [B0030] Ar  [J23110] Ar   **Electrophoresis of** PCR products:  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar  [B0030] Ar  [J23110] Ar   **Cultivation of**  [B0030] Ar  [J23110] Ar   **Ligation of**  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar |
| **10**  **Electrophoresis of**  [B0030] Ar   **Cultivation of**  [B0030] Ar    **Plasmid purification of**  [J23110] Ar  [B0030] Ar  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar   **Digestion of**   [B0030] Ar (EcoRI and SpeI)  [J23110] Ar (EcoRI and SpeI)  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)  GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI)   **Electrophoresis of** digested products:  [B0030] Ar (EcoRI and SpeI)  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)  GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI) | **11**  **Electrophoresis of** digested products:  [J23110] Ar (EcoRI and SpeI)   **Ligation of**  RFP[B0030+E1010+J61048+pSB1C3] Cr  GFP[B0030+E0040+J61048+pSB1K3] Kr   **Transformation off**  RFP[B0030+E1010+J61048+pSB1C3] Cr  GFP[B0030+E0040+J61048+pSB1K3] Kr   **PCR of**  RFP[B0030+E1010+J61048+pSB1C3] Cr  GFP[B0030+E0040+J61048+pSB1K3] Kr | **12**  **PCR of** ligated products:  RFP[B0030+E1010+J61048+pSB1C3] Cr   **Cultivation of** PCR products:  RFP[B0030+E1010+J61048+pSB1C3] Cr   **Plasmid purification of**  RFP[B0030+E1010+J61048+pSB1C3] Cr   **Digestion of**   RFP[B0030+E1010+J61048] Cr (EcoRI and SpeI)  RFP[B0030+E1010+J61048] Cr (EcoRI and PstI)   **Electrophoresis of**  digested products:  RFP[B0030+E1010+J61048] Cr (EcoRI and SpeI)  RFP[B0030+E1010+J61048] Cr (EcoRI and PstI)  PCR products: RFP[B0030+E1010+J61048+pSB1C3] Cr | **13**  **Ligation of**  [J23110+B0034+pSB1K3] Kr | **14**  **Cultivation of**  OmpA[J23101+B0034+OmpA+pSB1A3] Ar   **Transformation off**  [J23110+B0034+pSB1K3] Kr   **Electrophoresis of** digested products:  GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)  GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI)  [pSB1K3] Kr (EcoRI and PstI) | **15**  **Digestion of**   GBP[B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)   **Ligation of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr | **16**  **Transformation off**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr   **Electrophoresis of**  GBP[B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)  GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI)   **Ligation of**  GBP[B0034+FadL-GBP+pSB1C3] Cr   **Transformation off**  [J23110+B0034+pSB1K3] Kr |
| **17**  **Ligation of**  [J23110+B0034+pSB1K3] Kr  GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr   **PCR of**  GBP[B0034+FadL-GBP+pSB1C3] Cr  GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr   **Electrophoresis of** PCR products:  GBP[B0034+FadL-GBP+pSB1C3] Cr  GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr | **18**  **Cultivation of**  GBP[B0034+FadL-GBP+pSB1C3] Cr   **Plasmid purification of**  GBP[B0034+FadL-GBP+pSB1C3] Cr   **Digestion of**   GBP[B0034+FadL-GBP] Cr (XbaI and PstI)  RFP[B0030+E1010+J61048] Cr (XbaI and PstI)  GFP[B0030+E0040+J61048] **Transformation off**  [J23110+B0034+pSB1K3] Kr  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr  GFP[B0030+E0040+J61048] | **19**  **Ligation of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr   **PCR of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr   **Electrophoresis of**  digested products:  RFP[B0030+E1010+J61048] Cr (XbaI and PstI)  GBP[B0034+FadL-GBP] Cr (XbaI and PstI)  PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr   **Transformation off**  scFv-B[scFv-B+pSB1K3] Kr  scFv-C[scFv-C+pSB1K3] Kr  GFP[B0030+E0040+J61048]   **Cultivation of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr | **20**  **Cultivation of**  OmpA[J23101+B0034+OmpA] Ar  scFv-B[scFv-B+pSB1K3] Kr  scFv-C[scFv-C+pSB1K3] Kr   **Plasmid purification of**  OmpA[J23101+B0034+OmpA] Ar  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr  scFv-B[scFv-B+pSB1K3] Kr  scFv-C[scFv-C+pSB1K3] Kr   **Digestion of**   OmpA[J23101+B0034+OmpA] Ar (EcoRI and Pst1)  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr (EcoRI and PstI)  OmpA[J23101+B0034+OmpA] Ar (NcoI and PstI)  scFv-B[scFv-B] Ar (EcoRI and PstI)  scFv-C[scFv-C] Ar (EcoRI and PstI)   **Ligation of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr **Transformation off**  [J23110+B0034+pSB1K3] Kr  [pSB3C5] 抗性   **Electrophoresis of**  OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI) | **21**  **Digestion of**   scFv-B[scFv-B] Ar (EcoRI and PstI)  scFv-C[scFv-C] Ar (EcoRI and PstI)   **Electrophoresis of**  OmpA[J23101+B0034+OmpA] Ar (NcoI and PstI)  scFv-B[scFv-B] Ar (EcoRI and PstI)  scFv-C[scFv-C] Ar (EcoRI and PstI)   **Ligation of**  scFv-B[scFv-B+pSB1C3] Cr  scFv-C[scFv-C+pSB1C3] Cr  scFv-B[scFv-B+pSB1K3] Kr  scFv-C[scFv-C+pSB1K3] Kr | **22**  **Cultivation of**  [pSB3C5] Cr  [J23110+B0034+pSB1K3] Kr   **Transformation off**  GBP[J23110+B0034+GBP] Ar   **PCR of**  scFv-B[scFv-B+pSB1C3] Cr  scFv-C[scFv-C+pSB1C3] Cr  scFv-B[scFv-B+pSB1K3] Kr  scFv-C[scFv-C+pSB1K3] Kr   **Electrophoresis of**  scFv-B[scFv-B+pSB1C3] Cr  scFv-C[scFv-C+pSB1C3] Cr  scFv-B[scFv-B+pSB1K3] Kr  scFv-C[scFv-C+pSB1K3] Kr   **Ligation of**  scFv-B[scFv-B+pSB1A3] Ar  scFv-C[scFv-C+pSB1A3] Ar   **Transform of**  scFv-B[scFv-B+pSB1A3] Ar  scFv-C[scFv-C+pSB1A3] Ar | **23**  **Plasmid purification of**  [pSB3C5] Cr  [J23110+B0034+pSB1K3] Kr   **Digestion of**   [pSB3C5] Cr (EcoRI and PstI)  [J23110+B0034+pSB1K3] Kr (EcoRI and PstI)   **PCR of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr   **Transformation off**  scFv-B[scFv-B+pSB1A3] Ar  scFv-C[scFv-C+pSB1A3] Ar |
| **24**  **Electrophoresis of** digested products:  [pSB3C5] Cr (EcoRI and PstI)  [J23110+B0034+pSB1K3] Kr (EcoRI and PstI)   **Cultivation of**  PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr  scFv-B[scFv-B+pSB1C3] Cr  scFv-C[scFv-C+pSB1A3] Ar   **Plasmid purification of**  scFv-B[scFv-B+pSB1A3] Ar  scFv-C[scFv-C+pSB1A3] Ar  GFP[B0030+E0040+J61048]   **Digestion of**   [J23101+B0034] Kr (EcoRI and SpeI)   **Ligation of**  OmpA[J23101+B0034+OmpA+pSB3C5] Cr   **PCR of**  GBP[J23110+B0034+GBP] Ar by two pArts   **Electrophoresis of**  GBP[J23110+B0034+GBP] Ar | **25**  **Transformation off**  OmpA[J23101+B0034+OmpA+pSB3C5] Cr   **Cultivation of**  GBP[J23110+B0034+FadL-GBP] Ar   **Digestion of**   GBP[B003+FadL-GBP] Cr (XbaI and PstI)  scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI)  scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI)  GFP[B0030+E0040+J61048] (XbaI and PstI)   **Electrophoresis of**  GFP[B0030+E0040+J61048]  scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI)  scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI)  GFP[B0030+E0040+J61048] (XbaI and PstI)   **Cultivation of**  scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI)  scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI)  GFP[B0030+E0040+J61048] (XbaI and PstI) | **26**  **Ligation of**  OmpA[J23110+B0034+OmpA+pSB1A3] Ar  GBP[J23110+B0034+GBP+pSB1A3] Ar   **Gel extraction of**  [J23110] Ar (SpeI and PstI)   **Plasmid purification of**  GBP[J23110+B0034+GBP] Ar by two pArts   **Digestion of**   GBP[J23110+B0034+GBP] Ar by two pArts (EcoRI and SpeI) | **27**  **PCR of**  [J23101+B0034+OmpA+pSB3C5]   **Electrophoresis of**  GBP[J23110+B0034+GBP] Ar by two pArts (EcoRI and SpeI)  GBP[B0034+FadL-GBP] Cr (XbaI and PstI)  [J23110] Ar (SpeI and PstI) | **28**  **Ligation of**  scFv-B[scFv-B+pSB1A3] Ar  scFv-C[scFv-C+pSB1A3] Ar  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] 抗性??  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar   **Transform of**  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048]  scFv-B[scFv-B+pSB1A3] Ar  scFv-C[scFv-C+pSB1A3] Ar   **PCR of**  [J23101+B003+OmpA+pSB3C5] + primer(EX-OmpA and OmpA-S)  OmpA[J23101+B0034+OmpA+pSB3C5] Cr    Plasmid purification in gel slices of PCR products:  OmpA[EX-OmpA-S]   **Gel extraction of**  OmpA[OmpA] (EcoRI and SpeI)   **Transformation off**  OmpA[J23101+B0034+OmpA+pSB3C5] | **29**  **Plasmid purification of**  [J23110] Ar  [pSB1C3]   **Digestion of**   [J23110] Ar (EcoRI and SpeI)  [pSB1C3] (EcoRI and SpeI)   **Ligation of**  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar  [J23110+B0034+pSB1K3] Kr   **Electrophoresis of**  [J23110] Ar (EcoRI and SpeI)  [pSB1C3] (EcoRI and SpeI)   **Transform of**  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar  [J23110+B0034+pSB1K3] Kr  GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr | **30**  **PCR of**  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar  [J23110+B0034+pSB1K3] Kr  GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr |
| **31**  **Cultivation of**  [J23110+B0034] Kr  GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar   **Plasmid purification of**  [J23110+B0034] Kr  GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar   **Digestion of**   [J23110+B0034] Kr (EcoRI and PstI)  GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar (EcoRI and PstI)  [J23110+B0034] (EcoRI and SpeI) Kr  [J23101+B0034] (EcoRI and SpeI) Kr  OmpA[OmpA] Cr (EcoRI and PstI)  [pSB1K3] (EcoRI and PstI)   **Electrophoresis of**  [J23110+B0034] Kr (EcoRI and PstI)  GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar (EcoRI and PstI)   **Ligation of**  OmpA[OmpA+pSB1C3] Cr |  |  |  |  |  |  |

|  |  |  |  |  |  |  |
| --- | --- | --- | --- | --- | --- | --- |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
|  | **1**  **Electrophoresis of**  [J23110+B0034] (EcoRI and SpeI) Kr  [J23101+B0034] (EcoRI and SpeI) Kr  OmpA[OmpA] (NcoI and PstI)  GFP[K1489002+E0040] Kr   **Digestion of**   [pSB1C3] (EcoRI and PstI)  [pSB1A3] (EcoRI and PstI)  OmpA[OmpA] (EcoRI and NcoI)  [pSB3C5] (EcoRI and PstI)   **Ligation of**  GFP[J23101+B0034+K1489002+E0040+pSB3C5] (Cr)  GFP[J23110+B0034+K1489002+E0040+pSB3C5] (Cr)   **Electrophoresis of** digested product:  [pSB1C3] (EcoRI and PstI)  [pSB1A3] (EcoRI and PstI)  OmpA[OmpA] (EcoRI and NcoI)   **Cultivation of**  BFP[OmpA+K592100]   **PCR of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr   **Electrophoresis of**  GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr | **2**  **PCR of**  OmpA[EX-OmpA-S+pSB1C3] Cr   **Electrophoresis of**  OmpA[EX-OmpA-S+pSB1C3] Cr  OmpA[OmpA] (EcoRI and NcoI)   **Ligation of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E1010+J61048+pSB1K3] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr   **Digestion of**   [pSB1A3] (EcoRI and NcoI)   **Electrophoresis of** digested products:  [pSB1A3] (EcoRI and NcoI) | **3**  **Ligation of**  OmpA[OmpA+psB1C3] Cr  [J23110+B0034+pSB1K3] Kr   **Transform of**  OmpA[OmpA+psB1C3] Cr  [J23110+B0034+pSB1K3] Kr   **Digestion of**   BFP[OmpA+ K592100] (NcoI and PstI)  OmpA[OmpA] (NcoI and PstI) | **4**  **Cultivation of**  [J23110+pSB1A3] Ar   **Transform of**  [pSB1C3]   **Digestion of**   [pSB1C3] (EcoRI and PstI)   **Ligation of**  OmpA[OmpA+pSB1C3] Cr  OmpA[J23101+B0034+OmpA+pSB3C5] Cr   **Transform of**  [B0034+pSB1A3] Ar | **5**  **Transform of**  OmpA[J23101+B0034+OmpA+pSB3C5] Cr   **Plasmid purification of**  [J23110+pSb1A3] Ar   **Digestion of**   [J23110+pSb1A3] Ar   **PCR of**  [J23110+pSb1A3] Ar  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr   **Transform of**  OmpA[OmpA+pSB1C3] Cr   **Electrophoresis of**  [pSB1C3]  [B0034+pSB1A3] Ar  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr | **6**  **Cultivation of**  [pSB1C3]   **Digestion of**   [pSB1C3] (EcoRI and SpeI) |
| **7**  **Gel extraction of**  OmpA[EX-OmpA-S] 抗性  [pSB1C3] (EcoRI and SpeI)   **Gel extraction of**  [pSB1C3] (EcoRI and SpeI)   **Transform of**  GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar  [pSB1A3]  OmpA[J23101+B0034+OmpA] Ar   **Ligation of**  OmpA[OmpA+pSB1C3] Cr   **Cultivation of**  [pSB1C3] | **8**  **Transform of**  OmpA[OmpA+pSB1C3] Cr  [pSB3K3] [ pSB6A1]   **Plasmid purification of**  [pSB1C3]   **Digestion of**   OmpA[EX-OmpA-S] (EcoRI and SpeI)   **Electrophoresis of**  [pSB1C3] (EcoRI and SpeI)   **Transform of**  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr | **9**  **Ligation of**  OmpA[OmpA+pSB1C3] Cr   **PCR of**  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr   **Cultivation of**  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr   **Plasmid purification of**  [pSB3K3]  [pSB6A1]   **Digestion of**   [pSB3K3] (EcoRI and PstI)  [pSB6A1] (EcoRI and PstI) | **10**  **Transform of**  [OmpA+pSB1C3] Cr   **Plasmid purification of**  [J23101+B0034+OmpA+K592100+pSB3C5] Cr   **Digestion of**   [J23101+B0034+OmpA+K592100] Cr (EcoRI and PstI)   **Electrophoresis of**  [pSB3K3] (EcoRI and PstI)  [pSB6A1] (EcoRI and PstI)  [pSB1A3] (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B] (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C] (EcoRI and PstI)   **Transform of**  [J23110+B0034] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3]  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3]   **Ligation of**  scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr   **Electrophoresis of**  [pSB3K3]  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr | **11**  **Transform of**  scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr  [J23110+B0034+pSB1C3] Cr   **Ligation of**  [J23110+B0034+pSB1K3] Kr  [J23110+B0034+pSB1C3] Cr   **PCR of**  OmpA[OmpA+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr   **Electrophoresis of**  OmpA[OmpA+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr   **Cultivation of**  OmpA[OmpA+pSB1C3] Cr | **12**  **PCR of**  [J23110+B0034+pSB1C3] Cr  [B0034+pSB1A3] Ar   **Plasmid purification of**  OmpA[OmpA+pSB1C3] Cr   **Ligation of**  [J23110+B0034+pSB1C3] Cr   **Transform of**  [J23110+B0034+pSB1C3] Cr | **13**  **Electrophoresis of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI) |
| **14**  **SDS-page of**  [pSB1C3]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)  [pSB3C5]  scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr  scFv-B[OmpA+scFv-B]  scFv-C[OmpA+scFv-C]   **Electrophoresis of**  [J23110]  [B0034+FadL-GBP+B0030+E1010+J61048] 抗性 (XbaI and PstI)   **Ligation of**  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3K3] Cr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Cr   **Digestion of**   OmpA[OmpA+pSB1C3] Cr (EcoRI and SpeI)  OmpA[OmpA+pSB1C3] Cr (EcoRI and PstI)   **Transform of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar   **Ligation of**  GBP[FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar   **Transform of**  GBP[FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar | **15** | **16**  **Transform of**  [pSB3C5] | **17**  **Cultivation of**  [pSB3C5] | **18**  **Plasmid purification of**  [pSB3C5]   **Digestion of**   [pSB3C5]( EcoRI and PstI)   **Electrophoresis of**  [pSB3C5]( EcoRI and PstI)   **Ligation of**  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr | **19**  **Transform of**  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr | **20**  **PCR of**  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr   **Electrophoresis of**  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr |
| **21** | **22** | **23**  **PCR of**  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr   **Cultivation of**  [pSB3C5]  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  BFP[OmpA+K592100]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1]  [psB6A1]  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar  [pSB1A3]  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr   **Ligation of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar | **24**  **Plasmid purification of**  [pSB3C5] | **25**  **Cultivation of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr   **Transform of**  [J23101+B0034+pSB1K3] Kr | **26** | **27**  **Digestion of**   OmpA[OmpA] Cr (EcoRI and PstI)   **Electrophoresis of**  GBP[FadL-GBP] 抗性 (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar (EcoRI and PstI)  OmpA[OmpA] Cr (EcoRI and PstI) |
| **28**  **Cultivation of**  BFP[J23101+B0034+OmpA+K592100+pSB1A3] Ar  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3C5] Cr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  [pSB6A1]  [pSB3C5]  [pSB1C3]  [pSB1A3]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr   **Electrophoresis of**  [J23101+B0034] Kr  GBP[FadL-GBP] 抗性 (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar (EcoRI and PstI)  OmpA[OmpA] Cr (EcoRI and PstI)  BFP[J23101+B0034+OmpA+K592100] Ar (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Cr (EcoRI and PstI)  [pSB6A1] (EcoRI and PstI)  OmpA[OmpA] Cr (XbaI and PstI)  [E0040] Ar (XbaI and PstI)   **Ligation of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr  [B0034+E0040+pSB1K3] Kr  GBP[FadL-GBP+pSB6A1] Ar  GBP[FadL-GBP+pSB1C3] Cr  GBP[FadL-GBP+pSB3C5] Cr   **Digestion of**   BFP[J23101+B0034+OmpA+K592100] Cr (XbaI and PstI)   **SDS-page of**  [pSB1A3]  [pSB1C3]  BFP[J23101+B0034+OmpA+K592100+pSB1A3] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr  [pSB3C5]  [pSB6A1]  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3C5] Cr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar   **Transform of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  GBP[FadL-GBP+pSB1C3] Cr  GBP[FadL-GBP+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr | **29**  **PCR of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  GBP[FadL-GBP+pSB1C3] Cr  GBP[FadL-GBP+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr   **Electrophoresis of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar | **30**  **Electrophoresis of**  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar |  |  |  |  |

|  |  |  |  |  |  |  |
| --- | --- | --- | --- | --- | --- | --- |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
|  |  |  | **1**  **Cultivation of**  [pSB6A1]  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar   **Transform of**  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr   **Digestion of**   GBP[]FadL-GBP] 抗性 (EcoRI and PstI) | **2**  **Digestion of**   [J23110] Ar (SpeI and PstI)    Plasmid purification in gel slices of  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar   **Digestion of**   scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)   **Electrophoresis of**  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) | **3**  **Gel extraction of**  [J23110] Ar (SpeI and PstI)  [pSB6A1] (EcoRI and PstI)   **Transform of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr   **Electrophoresis of**  GBP[FadL-GBP] (EcoRI and Pstl)   **Ligation of**  GBP[FadL-GBP+pSB3C5] Cr  GBP[FadL-GBP+pSB1C3] Cr  GBP[FadL-GBP+pSB6A1] Ar | **4**  **Transform of**  GBP[FadL-GBP+pSB3C5] Cr  GBP[FadL-GBP+pSB1C3] Cr  GBP[FadL-GBP+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr   **Electrophoresis of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr   **Cultivation of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr  [J23101+B0034+pSB1K3] Kr |
| **5**  **Plasmid purification of**  [J23101+B0034+pSB1K3] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr | **6**  **Cultivation of**  [pSB1C3]  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  [pSB6A1]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  [pSB1A3]  GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar | **7**  **Digestion of**   [J23101+B0034+pSB1K3] Kr (EcoRI and PstI)   **SDS-page of**  [pSB1C3]  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  [pSB6A1]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  [pSB1A3]  GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar   **Ligation of**  [J23110+B0034] Ar   **Electrophoresis of**  [J23101+B0034+pSB1K3] Kr (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar | **8**  **Plasmid purification of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar   **Transform of**  [J23110+B0034] Ar  BFP[J23101+B0034+OmpA+K592100]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100]   **Electrophoresis of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) | **9**  **Electrophoresis of**  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+scFv-B+B0034+E0040+J61048+pSB1C3] Cr  [pSB3C5]  BFP[J23101+B0034+OmpA+scFv-B+B0030+e0040+J61048+pSB6A1] Ar   **Cultivation of**  scFv-B[J23110+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar | **10**  **Plasmid purification of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar | **11**  **PCR of**  [J23110+B0034] Ar   **Electrophoresis of**  [J23110+B0034] Ar |
| **12**  **Cultivation of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  [J23110+B0034] Ar   **Plasmid purification of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar   **Digestion of**   scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) | **13**  **Plasmid purification of**  [J23110+B0034] Ar   **Transform of**  [R0010]   **Electrophoresis of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) | **14**  **Digestion of**   [J23110+B0034] Ar (EcoRI and PstI)  GBP[FadL-GBP] Kr (XbaI and PstI)  OmpA[OmpA] Cr (EcoRI and SpeI)    **Electrophoresis of**  [J23110+B0034] Ar (EcoRI and PstI)  GBP[FadL-GBP] Kr (XbaI and PstI)  OmpA[OmpA] Cr (EcoRI and SpeI)   **Transform of**  [R0010] Cr   **Cultivation of**  [pSB1C3]  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr | **15**  **Digestion of**   [pSB6A1]  [pSB3C5]  Anti-Her 2[Herceptin+E1010]  [J23101+B0034+OmpA+scFv-C]  [J23101+B0034+OmpA+scFv-B]   **Cultivation of**  [pSB6A1]  [R0010]   **PCR of**  OmpA[OmpA-linker] (EcoRI and PstI)   **Transform of**  [J23110+B0034] Kr  BFP[K592100+J61048] Kr | **16**  **Gel extraction of**  OmpA[OmpA-linker] (EcoRI and PstI)   **Ligation of**  OmpA[J23101+B0034+OmpA+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr   **Digestion of**   OmpA[OmpA-linker] (EcoRI and PstI)   **Transform of**  OmpA[J23101+B0034+OmpA+pSB3C5] Cr | **17**  **Electrophoresis of**  OmpA[OmpA-linker] (EcoRI and PstI)  Anti-Her 2[Herceptin+E1010] (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) | **18**  **Ligation of**  OmpA[OmpA-linker+pSB1C3] Cr  Anti-Her 2[Herceptin+E1010+pSB1C3] Cr |
| **19**  **Transform of**  OmpA[OmpA-linker+pSB1C3] Cr  Anti-Her 2[Herceptin+E1010+pSB1C3] Cr | **20** | **21** | **22** | **23** | **24**  **Transform of**  OmpA[OmpA-linker+pSB1C3] Cr  Anti-Her 2[Herceptin+E1010+pSB1C3] Cr   **PCR of**  OmpA[OmpA-linker+pSB1C3] Cr  Anti-Her 2[Herceptin+E1010+pSB1C3] Cr   **Electrophoresis of**  OmpA[OmpA-linker+pSB1C3] Cr  Anti-Her 2[Herceptin+E1010+pSB1C3] Cr | **25**  **Cultivation of**  [R0010] Cr  BFP[K592100+J61048] Kr   **PCR of**  OmpA[OmpA-linker+pSB1C3] Cr  Anti-Her 2[Herceptin+E1010+pSB1C3] Cr   **Electrophoresis of**  OmpA[OmpA-linker+pSB1C3] Cr  Anti-Her 2[Herceptin+E1010+pSB1C3] Cr |
| **26**  **Plasmid purification of**  [R0010] Cr  BFP[K592100+J61048] Kr   **Digestion of**   [R0010] Cr (EcoRI and SpeI)  BFP[K592100+J61048] Kr (EcoRI and PstI)   **Electrophoresis of**  [R0010] Cr  BFP[K592100+J61048] Kr   **Ligation of**  [R0010+B0034+pSB1K3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr  BFP[B0030+K592100+J61048+pSB1C3] Cr   **Transform of**  scFv-C[J23101+B0034+OmpA+scFv-C] Ar   **Ligation of**  OmpA[OmpA-linker+pSB1C3] Cr   **PCR of**  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048]   **Cultivation of**  [pSB1C3]  GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr  BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr  GBP[FadL-GBP+pSB6A1] Ar | **27**  **PCR of**  GBP[FadL-GBP]  Anti-Her 2[Herceptin+E1010]   **Gel extraction of**  GBP[FadL-GBP]  Anti-Her 2[Herceptin+E1010]   **Transform of**  [R0010+B0034+pSB1K3]  GBP[R0010+B0034+FadL-GBP+B0030+J61048+pSB1K3]  BFP[B0030+K592100+J61048+pSB1C3] Cr   **PCR of**  GBP[J23110+B0034+FadL-GBP+E0040+J61048] Cr   **Gel extraction of**  GBP[J23110+B0034+FadL-GBP+E0040+J61048] Cr   **SDS-page of**  [pSB1C3]  GFP[J23101+B0034+E0040+J61048+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr  BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr  GBP[J23110+B0034+FadL-GBP+E0040+J61048+pSB1C3] Cr  GBP[FadL-GBP+pSB6A1] Ar  [pSB6A1] | **28**  **Ligation of**  [R0010+B0034+pSB1K3]  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1K3] Kr  BFP[B0030+K592100+J61048+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar   **PCR of**  BFP[B0030+K592100+J61048+pSB1C3] Cr  [R0010+B0034+pSB1K3]  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1K3] Kr  OmpA[J23110+B0034+OmpA+pSB3C5] Cr   **Digestion of**   GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048] (XbaI and PstI)   **Electrophoresis of**  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048] (XbaI and PstI)   **SDS-page of**  [pSB1C3]  BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr  GFP[J23101+B0034+E0040+J61048+pSB1C3] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr  [pSB6A1]  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar   **Transform of**  OmpA[OmpA-linker] Cr | **29**  **Electrophoresis of**  OmpA[J23101+B0034+OmpA+pSB3C5]   **Transform of**  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar   **PCR of**  BFP[B0034+K592100+J61048+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]   **Electrophoresis of**  BFP[B0034+K592100+J61048+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]   **Digestion of**   Anti-Her 2[Herceptin-own] (EcoRI and PstI)  Anti-Her 2[Herceptin-E1010] (EcoRI and PstI)  GBP[GBP] (EcoRI and PstI)   **Ligation of**  [R0010+B0034+pSB1K3] Kr  BFP[B0030+K592100+J61048] Cr  GBP[R0010+B0034+FadL-GBP+B0034+E0040+J61048] Ar  GFP[R0010+B0030+E0040+J61048] Ar   **Electrophoresis of**  Anti-Her 2[Herceptin-own] (EcoRI and PstI)  Anti-Her 2[Herceptin-E1010] (EcoRI and PstI)  GBP[GBP] (EcoRI and PstI)  [R0010+B0034+pSB1K3] Kr  BFP[B0030+K592100+J61048] Cr  GBP[R0010+B0034+FadL-GBP+B0034+E0040+J61048] Ar  GFP[R0010+B0030+E0040+J61048] Ar | **30**  **Cultivation of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr   **Ligation of**  GBP[FadL-GBP+pSB1C3]Cr   **Digestion of**   Anti-Her 2[Herceptin own] (NcoI and PstI)  [pSB1A3] (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)   **Cultivation of**  scFv-C[J23101+B0034+OmpA+scFv-C] Ar  [J23101+B0034] Kr   **Digestion of**   scFv-B[scFv-B] Kr (NcoI and PstI)  scFv-C[scFv-C] Kr (NcoI and PstI)  OmpA[OmpA] Cr (EcoRI and PstI)   **Electrophoresis of**  Anti-Her 2[Herceptin own] (NcoI and PstI)  [pSB1A3] (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)   **Plasmid purification of**  scFv-C[J23101+B0034+OmpA+scFv-C] Ar  [J231-1+B0034] Kr   **Digestion of**   scFv-C[J23101+B0034+OmpA+scFv-C] Ar (EcoRI and SpeI)  [J231-1+B0034] Kr (EcoRI and SpeI)  Anti-Her 2[Herceptin-Own] (NcoI and PstI) | **31**  **Electrophoresis of**  scFv-C[J23101+B0034+OmpA+scFv-C] Ar (EcoRI and SpeI)  [J231-1+B0034] Kr (EcoRI and SpeI)  Anti-Her 2[Herceptin-Own] (NcoI and PstI)   **Ligation of**  Anti-Her 2[J2310+B0034+OmpA+Herceptin-Own+pSB1A3] Ar   **Transform of**  Anti-Her 2[J2310+B0034+OmpA+Herceptin-Own+pSB1A3] Ar   **Electrophoresis of**  [pSB1C3] (XbaI and PstI)  OmpA[OmpA] Cr (EcoRI and PstI)  scFv-B[scFv-B] Kr (NcoI and PstI)  scFv-C[scFv-C] Kr (NcoI and PstI) |  |

|  |  |  |  |  |  |  |
| --- | --- | --- | --- | --- | --- | --- |
| Sun. | Mon. | Tue. | Wed. | Thu. | Fri. | Set. |
|  |  |  |  |  |  | **1**   **Digestion of**   Anti-Her 2[Herceptin-Web] (NcoI and PstI)   **Electrophoresis of**  Anti-Her 2[Herceptin-Web] (NcoI and PstI) |
| **2**  **Electrophoresis of**  [pSB1C3] (XbaI and PstI)  OmpA[OmpA] (EcoRI and PstI)  scFv-B[scFv-B] Kr (NcoI and PstI)  scFv-C[scFv-C] Kr (NcoI and PstI)  OmpA[OmpA] (NcoI and PstI)   **Digestion of**   BFP[OmpA+K592100] Cr (EcoRI and NcoI) | **3**  **Transform of** [R0010+B0034+pSB1K3] Kr  GBP[FadL-GBP+pSB1C3] Cr  BFP[B0030+K592100+J61048] Cr   **Digestion of**   Anti-Her 2[Herceptin-Web] (NcoI and PstI)   **Ligation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar | **4**  **Digestion of**   BFP[OmpA+K592100] Cr (EcoRI and NcoI)   **PCR of**  [R0010+B0034+pSB1K3] Kr  BFP[B0030+K592100+J61048] Cr   **Transform of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar | **5**  **Digestion of**   [pSB6A1] (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)   **PCR of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **Electrophoresis of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **Cultivation of**  BFP[B0030+K592100+J61048] Cr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]   **PCR of**  GBP[FadL-GBP] Cr  BFP[OmpA+K592100] Cr  scFv-C[J23101+B0034+OmpA+scFv-C] Ar  scFv-C[scFv-C+pSB1C3] Cr  scFv-B[scFv-B+oSB1C3] Cr   **Digestion of**   scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr (EcoRI and PstI)   **Transform of**  GBP[J23101+B0034+FadL-GBP]Cr  scFv-B[scFv-B+pSB1C3] Cr  scFv-C[scFv-C+pSB1C3] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr   **Gel extraction of**  [pSB6A1] (EcoRI and PstI)  [OmpA] (NcoI and PstI)  [pSB1C3] (EcoRI and PstI) | **6**  **Cultivation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **Ligation of**  Anti-Her 2[OmpA+Herceptin-Web] Cr  Anti-Her 2[OmpA+Herceptin-Own] Cr  scFv-C[OmpA+scFv-C] Cr  scFv-B[OmpA+scFv-B] Cr  scFv-C[J23101+B0034+OmpA+scFv-C] Cr  scFv-B[J23101+B0034+OmpA+scFv-B] Cr   **Transform of**  Anti-Her 2[OmpA+Herceptin-Web] Cr  Anti-Her 2[OmpA+Herceptin-Own] Cr  scFv-C[OmpA+scFv-C] Cr  scFv-B[OmpA+scFv-B] Cr  scFv-C[J23101+B0034+OmpA+scFv-C] Cr  scFv-B[J23101+B0034+OmpA+scFv-B] Cr  [pSB1C3]  [pSB1A3]  [pSB1K3]   **Plasmid purification of**  BFP[B0030+K592100+J61048]   **Digestion of**   BFP[B0030+K592100+J61048] (EcoRI and PstI)   **Cultivation of**  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr   **Ligation of**  [R0010+B0034] Kr  GBP[GBP[J23101+B0034+FadL-GBP]+pSB1C3] Cr   **Digestion of**   [pSB1C3] (XbaI and PstI)   **Electrophoresis of**  GBP[J23101+B0034+FadL-GBP] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040] Cr  scFv-C[scFv-C] Cr  scFv-B[scFv-B] Cr   **Plasmid purification of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **Digestion of**   Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI)   **Ligation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar   **Electrophoresis of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar  [B0034]  [pSB1C3]   **Cultivation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **Transform of**  GBP[J23101+B0034+FadL-GBP] Ar   **Electrophoresis of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040] Cr  [pSB1C3] (XbaI and PstI)  [pSB6A1] (XbaI and PstI)  [pSB1C3] (XbaI and PstI)  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **Cultivation of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr  GFP[J23101+B0034+E0040+J61048] Ar   **SDS-page of**  GFP[J23101+B0034+E0040+J61048] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Ar  [pSB6A1]  GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Ar   **Digestion of**   [pSB1C3] (XbaI and PstI)  Anti-Her 2[Herceptin-Web] (NcoI and PstI)  Anti-Her 2[Herceptin-Own] (NcoI and PstI)   **Ligation of**  [pSB1C3] (XbaI and PstI)  Anti-Her 2[Herceptin-Web] Cr  Anti-Her 2[Herceptin-Own] Cr  BFP[J23101+B0034+OmpA+K592100] Cr | **7**  **Plasmid purification of**  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr   **Digestion of**   GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr   **PCR of**  scFv-B[J23101+B0034+OmpA+scFv-B] Cr  scFv-C[J23101+B0034+OmpA+scFv-C] Cr   **Electrophoresis of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI)   **Cultivation of**  scFv-B[J23101+B0034+OmpA+scFv-B] Cr  scFv-C[J23101+B0034+OmpA+scFv-C] Cr   **Transform of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **Plasmid purification of**  GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr   **Digestion of**   GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr (EcoRI and PstI)   **PCR of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar   **Electrophoresis of**  scFv-B[J23101+B0034+OmpA+scFv-B] Cr  scFv-C[J23101+B0034+OmpA+scFv-C] Cr  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar  OmpA[OmpA] (NcoI and PstI)  [pSB1C3] (EcoRI and PstI)   **Ligation of**  scFv-C[OmpA+scFv-C] Cr  scFv-B[OmpA+scFv-B] Cr  Anti-Her 2[Herceptin-Own] Cr  Anti-Her 2[Herceptin-Web] Cr  Anti-Her 2[OmpA+Herceptin-Own] Cr  Anti-Her 2[OmpA+Herceptin-Web] C   **Electrophoresis of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI)  Anti-Her 2[Herceptin-Web] (NcoI and PstI)  Anti-Her 2[Herceptin-Own] (NcoI and PstI)   **Transform of**  scFv-C[OmpA+scFv-C] Cr  scFv-B[OmpA+scFv-B] Cr  Anti-Her 2[Herceptin-Own] Cr  Anti-Her 2[Herceptin-Web] Cr  Anti-Her 2[OmpA+Herceptin-Own] Cr  Anti-Her 2[OmpA+Herceptin-Web] C  BFP[J23101+B0034+OmpA+K592100] Cr | **8**  **Plasmid purification of**  scFv-C[J23101+OmpA+scFv-C] Cr  scFv-B[J23101+OmpA+scFv-B] Cr   **Digestion of**   scFv-C[J23101+OmpA+scFv-C] Cr (EcoRI and PstI)  scFv-B[J23101+OmpA+scFv-B] Cr (EcoRI and PstI)   **PCR of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar   **Cultivation of**  [pSB1K3]  [pSB1C3]  [pSB1A3]   **PCR of**  BFP[J23101+B0034+OmpA+K592100] Cr   **Transform of**  scFv-C[OmpA+scFv-C] Cr   **Electrophoresis of**  BFP[J23101+B0034+OmpA+K592100] Cr  scFv-C[J23101+OmpA+scFv-C] Cr (EcoRI and PstI)  scFv-B[J23101+OmpA+scFv-B] Cr (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)   **Digestion of**   Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and SpeI)  OmpA[OmpA] Cr NcoI and PstI  OmpA[OmpA+K592100] Cr NcoI and PstI   **Ligation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr   **Digestion of**   Anti-Her 2[Herceptin-Own] (EcoRI and PstI)  Anti-Her 2[Herceptin-Web] (EcoRI and PstI)   **Electrophoresis of**  BFP[J23101+B0034+OmpA+K592100] Cr |
| **9**  **Cultivation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar   **Plasmid purification of**  [pSB1C3]  [pSB1K3]  [pSB1A3]   **Digestion of**   [pSB1C3] (EcoRI and PstI)  [pSB1K3] (EcoRI and PstI)  [pSB1A3] (EcoRI and PstI)   **Transform of**  Anti-Her 2[Herceptin-Own] Cr  Anti-Her 2[Herceptin-Web] Cr  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr  [R0010+B0034] Cr  BFP[J23101+B0034+OmpA+K592100] Cr   **Electrophoresis of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr (EcoRI and SpeI)  OmpA[OmpA] Cr (NcoI and PstI)   **Digestion of**   [pSB1A3] (EcoRI and PstI)  [pSB1K3] (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)  RFP[B0030+E1010+J61048] Cr (XbaI and PstI)   **Electrophoresis of**  scFv-C[OmpA+scFv-C] Kr   **PCR of**  GBP[GBp+pSB1C3] Cr   **Digestion of**   BFP[B0030+K592100+J61048] Cr (XbaI and PstI) | **10**  **Electrophoresis of**  GBP[GBp+pSB1C3] Cr  [pSB1A3] (EcoRI and PstI)  [pSB1K3] (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)   **Ligation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web+B0030+E1010+J61048] Kr   **Cultivation of**  [R0010+B0034] Cr  BFP[B0030+K592100+J61048] Cr  scFv-C[OmpA+scFv-C] Kr   **Digestion of**   [pSB1C3] (EcoRI and Pstl)   **Transform of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr   **Gel extraction of**  [pSB1C3] (EcoRI and PstI)   **PCR of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web]  Anti-Her 2[Herceptin-Web]  Anti-Her 2[Herceptin-Own] | **11**  **Plasmid purification of**  [R0010+B0034] Cr  scFv-C[OmpA+scFv-C] Kr  BFP[OmpA+K592100] Cr    **Gel extraction of**  [pSB1C3] (EcoRI and PstI)   ******Digestion of******   [R0010+B0034] Cr (EcoRI and SpeI)  scFv-C[OmpA+scFv-C] Kr (EcoRI and PstI)  BFP[OmpA+K592100] Cr (EcoRI and PstI)  GBP[J23101+B0034+FadL-GBP] Cr (EcoRI and SpeI)   **Ligation of**  Anti-Her 2[Herceptin-Web] Cr   **PCR of**  scFv-B[OmpA+scFv-B] Kr   **Electrophoresis of**  [R0010+B0034] Cr (EcoRI and SpeI)  scFv-C[OmpA+scFv-C] Kr (EcoRI and PstI)  BFP[OmpA+K592100] Cr (EcoRI and PstI)  GBP[J23101+B0034+FadL-GBP] Cr (EcoRI and SpeI)   **Ligation of**  GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar   **Electrophoresis of**  scFv-B[OmpA+scFv-B] Kr   **PCR of**  scFv-C[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr   **Transform of**  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr | **12**  **Electrophoresis of**  scFv-B[OmpA+scFv-B] Kr   **Transform of**  Anti-Her 2[Herceptin-Web] Cr  Anti-Her 2[Herceptin-Own] Cr  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar   **PCR of**  scFv-C[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar   **Digestion of**   [pSB1C3] (EcoRI and PstI)   **Electrophoresis of**  scFv-C[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar | **13**  **Transform of**  GBP[J231010+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar   **Cultivation of**  [pSB1C3]  scFv-B[OmpA-scFv-B] Kr   **Plasmid purification of**  [pSB1C3]  scFv-B[OmpA-scFv-B] Kr   **Digestion of**   [pSB1C3] (EcoRI and PstI)  scFv-B[OmpA-scFv-B] Kr (EcoRI and PstI)   **Electrophoresis of**  [pSB1C3] (EcoRI and PstI)   **Ligation of**  OmpA[R0010+B0034+OmpA+pSB1A3] Ar  GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar  scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar   **Electrophoresis of**  [pSB1C3] (EcoRI and PstI)  scFv-B[OmpA-scFv-B] Kr (EcoRI and PstI) | **14**  **Transform of**  OmpA[R0010+B0034+OmpA+pSB1A3] Ar  GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar | **15**  **PCR of**  OmpA[R0010+B0034+OmpA+pSB1A3] Ar  GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar  scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar   **Ligation of**  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr  BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr   **PCR of**  GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr   **Transform of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr   **Electrophoresis of**  GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr   **Electrophoresis of**  GBP[FadL-GBP] (EcoRI and PstI) |
| **16**  **PCR of**  GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Kr   **Cultivation of**  OmpA[R0010+B0034+OmpA]   **Electrophoresis of**  GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Kr | **17**  **Ligation of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar   **Transform of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar | **18**  **Plasmid purification of**  OmpA[R0010+B0034+OmpA+pSB1A3] Ar  GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar   **Digestion of**   BFP[OmpA+K592100] Cr (EcoRI and NcoI)  Anti-Her 2[Herceptin-Own] (NcoI and SpeI)  Anti-Her 2[Herceptin-Web] (NcoI and SpeI)  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar (EcoRI and SpeI)  BFP[B0030+K592100+J61048] Cr (XbaI and PstI)  OmpA[R0010+B0034+OmpA+pSB1A3] Ar (EcoRI and PstI)  GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)   **Ligation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar   **Transform of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar  GBP[J23101+B0034+fadL-GBP+B0030+E0040+J61048] Cr  OmpA[R0010+B0034+OmpA+pSB1C3] Cr  GBP[R0010+B0034+FadL-GBP+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048]   **Ligation of**  scFv-B[R0010+B0034+OmpA+scFv-B]  scFv-C[R0010+B0034+OmpA+scFv-C]  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own]  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web]  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]   **PCR of**  Anti-Her 2[Herceptin-Web]  scFv-B[J23101+B0034+OmpA+scFv-B]  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr  Anti-Her 2[Herceptin-Own]  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar | **19**  **Electrophoresis of**  Anti-Her 2[Herceptin-Web]  scFv-B[J23101+B0034+OmpA+scFv-B]  scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr  Anti-Her 2[Herceptin-Own]  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar  scFv-C[OmpA+scFv-C]  Anti-Her 2[Herceptin-Own] (NcoI and SpeI)  Anti-Her 2[Herceptin-Web] (NcoI and SpeI)  Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar (EcoRI and SpeI)  BFP[B0030+K592100+J61048] Cr (XbaI and PstI)   **Transform of**  GBP[R0010+B034+FadL-GBP] Ar  GBP[J23101+B0034+FadL-GBP] Ar  GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr  **PCR of**  scFv-B[J23110+B0034+OmpA+scFv-B] | **20**  **PCR of**  GBP[FadL-GBP] Kr  OmpA[OmpA] Cr   **Gel extraction of**  OmpA[OmpA] (EcoRI and SpeI)   **Plasmid purification of**  Anti-Her 2[Herceptin-Own] Cr  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar   **PCR of**  GBP[FadL-GBP] Kr  OmpA[R0010+B0034+OmpA+pSB1C3] Cr  GBP[R0010+B0034+GBP+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr  GBP[R0010+B0034+GBP+B0030+E0040+J61048]  scFv-B[R0010+B0034+OmpA+scFv-B]  scFv-C[R0010+B0034+OmpA+scFv-C]  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own]  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web]   **Digestion of**   scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)  Anti-Her 2[Herceptin-Own] Cr   **Electrophoresis of**  OmpA[R0010+B0034+OmpA+pSB1C3] Cr  GBP[R0010+B0034+GBP+pSB1C3] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr  GBP[R0010+B0034+GBP+B0030+E0040+J61048]   **Ligation of**  scFv-B[R0010+B0034+OmpA+scFv-B+pSB1K3] Kr  scFv-C[R0010+B0034+OmpA+scFv-C+pSB1K3] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+pSB1K3] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web+pSB1K3] Kr  GBP[R0010+B0034+GBP+B0030+E0040+J61048pSB1A3] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048+pSB1K3] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+B0030+E1010+J61048+pSB1K3] Kr  **PCR of**  GBP[R0010+B0034+FadL-GBP]  GBP[J23110+B0034+FadL-GBP+B0030+E0040] | **21**   **Electrophoresis of**  [pSB1K3] (EcoRI and PstI)  [R0010] Cr (EcoRI and SpeI)  [J23110] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048] Ar (EcoRI and SpeI)  BFP[B0034+K592100+J61048] Cr (XbaI and PstI)  RFP[B0034+E1010+J61048] Cr (XbaI and PstI)  GBP[B0034+FadL-GBP+B0030+J61048] Cr (XbaI and PstI)  **Ligation of**  BFP[J23110+B0034+K592100+J61048+pSB1K3] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+K592100+E1010+J61048pSB1K3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3]  **Transform of**  scFv-B[R0010+B0034+OmpA+scFv-B+pSB1K3] Kr  scFv-C[R0010+B0034+OmpA+scFv-C+pSB1K3] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+pSB1K3] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web+pSB1K3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048pSB1A3] Ar  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048+pSB1K3] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+B0030+E1010+J61048+pSB1K3] Kr   **PCR of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own]  GBP[J23101+B0034+FadL-GBP]   **Electrophoresis of**  GBP[R0010+B0034+FadL-GBP] Ar  GBP[J23110+B0034+FadL-GBP+B0030+E0040] | **22**   **Digestion of**  [pSB1C3] (XbaI and SpeI)   **PCR of**  scFv-B[R0010+B0034+OmpA+scFv-B] Kr  scFv-C[R0010+B0034+OmpA+scFv-C] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-web] Kr  Anti-Her 2[R0010+B0034+OmpA+Herceptin-own] Ar  GBP[FadL-GBP+B0030+E0040+J61048] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048+pSB1K3] Kr  BFP[J23110+B0034+K592100+J61048+pSB1K3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040] Kr   **Cultivation of**  [pSB6A1]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048]   **Electrophoresis of**  GBP[R0010+B0034+FadL-GBP] Ar  GBP[FadL-GBP] (XbaI and PstI)  OmpA[OmpA] Cr (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)   **Digestion of**  [pSB1C3] (XbaI and PstI)   **Ligation of**  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048+pSB1K3]  GBP[R0010+B0034+FadL-GBp+B0030+E0040+J61048] Cr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] Ar  Anti-Her 2[R0010+B0034+Herceptin-own+B0030+E1010+J61048]  Anti-Her 2[R0010+B0034+Herceptin-own+B0030+E0040+J61048]  Anti-Her 2[R0010+B0034+Herceptin-own+B0030+K592100+J61048]  scFv-C[R0010+B0034+OmpA+scFv-C]   **Cultivation of**  GBP[R0010+B0034+FadL-GBp+B0030+E0040+J61048] Cr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] Ar |
| **23**   **Transform of**  GBP[FadL-GBP] Ar  OmpA[OmpA] Cr   **Plasmid purification of**  Anti-Her 2[R0010+B0034+Herceptin-web+B0030+E1010+J61048] Kr  Anti-Her 2[R0010+B0034+Herceptin-own+B0030+E1010+J61048] Kr  BFP[J23101+B0034+K592100+J61048] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr  GBP[R0010+B0034+FadL-GBP]  Anti-Her 2[R0010+B0034+Herceptin-own]   **Transform of**  Anti-Her 2[R0010+B0034+Herceptin-web+B0030+E1010+J61048] Kr  Anti-Her 2[R0010+B0034+Herceptin-own+B0030+E1010+J61048] Kr  BFP[J23101+B0034+K592100+J61048] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr   **Cultivation of**  GBP[R0010+B0034+FadL-GBP]  GBP[J23110+B0034+fadL-GBP+B0030+E0040]  scFv-B[R0010+B0034+OmpA+scFv-B] Kr   **Electrophoresis of**  scFv-B[J23101+B0034+OmpA+scFv-B] (EcoRI and SpeI)  GBP[B0034+FadL-GBP+B0030+E0040+J61048] (XbaI and PstI)  RFP[B0030+E1010+J61048] (XbaI and PstI)  BFP[J23101+B0034+K592100+J61048]  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr  Anti-Her 2[R0010+B0034+Herceptin-web] (EcoRI and PstI)  Anti-Her 2[R0010+B0034+Herceptin-own] (EcoRI and PstI)  GBP[R0010+B0034+FadL-GBP+B0030+E0040] (EcoRI and PstI)  GBP[R0010+B0034FadL-GBP] (EcoRI and PstI)  [pSB1C3] (XbaI and PstI) | **24**   **Ligation of**  scFv-B[OmpA+scFv-B] Kr  scFv-C[OmpA+scFv-C] Kr  Anti-Her 2[OmpA+Herceptin-own] Kr  Anti-Her 2[OmpA+Herceptin-web] Kr   **Digestion of**  Anti-Her 2[Herceptin-own] (NcoI and PstI)   **Electrophoresis of**  GBP[FadL-GBP] Cr (EcoRI and PstI)  GBP[J23110+B0034+FadL-GBP] (EcoRI and SpeI)  Anti-Her 2[R0010+B0034+OmpA+Herceptin-own] (EcoRI and SpeI)  GBP[J23110+B0034+FadL-GBP] (EcoRI and SpeI)  GBP[FadL-GBP] (EcoRI and PstI)   **Plasmid purification of**  GBP[R0010+B0034+FadL-GBP] Ar  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] Cr   **Digestion of**  GBP[R0010+B0034+FadL-GBP] Ar (EcoRI and SpeI)  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] Cr (EcoRI and PstI)   **Transform of**  GBP[FadL-GBP] Cr  OmpA[OmpA] Cr   **PCR of**  scFv-B[J23110+B0034+scFv-B+B0030+E1010+J61048+pSB1K3] Kr  Anti-Her 2[J23110+B0034+Herceptin-own+E0040+J61048+pSB1K3] Kr  scFv-C[R0010+B0034+OmpA+scFv-C+pSB1K3] Kr  Anti-Her 2[J23110+B0034+Herceptin-own+E1010+J61048+pSB1K3] Kr  Anti-Her 2[J23110+B0034+Herceptin-own+K592100+J61048+pSB1K3] Kr  scFv-B[J23110+B0034+Herceptin-own+K592100+J61048+pSB1K3] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+J61048] Kr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+J61048] Kr   **Transform of**  scFv-B[OmpA+scFv-B] Kr  scFv-C[OmpA+scFv-C] Kr  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+J61048] Kr  Anti-Her 2[OmpA+Herceptin-web] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr  scFv-B[J23110+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr  Anti-Her 2[OmpA+Herceptin-own] Kr  scFv-C[J23110+B0034+OmpA+scFv-C+B0030+E0040+J61048]  scFv-B[J23110+B0034+OmpA+scFv-B+B0030+E0040+J61048]   **SDS-page of**  scFv-C[J23110+B0034+OmpA+scFv-C+B0030+E0040+J61048]  scFv-B[J23110+B0034+OmpA+scFv-B+B0030+E0040+J61048]  GFP[J23110+B0034+E0040+J61048  GBP[R0010+B0034+FadL-GBP]  [pSB1C3]  OmpA[R0010+B0034+OmpA+J61048]  GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048]   **PCR of**  GBP[FadL-GBP] Cr  OmpA[OmpA]   **Ligation of**  BFP[J23101+B0034+OmpA+K592100] Cr  scFv-B[J23101+B0034+OmpA+scFv-B+] Cr | **25**  **Digestion of**  [pSB3C5] (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C] Ar (EcoRI and PstI)  scFv-B[J23101+B0034+OmpA+scFv-B+J61048] Cr (EcoRI and PstI)  scFv-B[R0010+B0034+OmpA+scFv-B] Kr (EcoRI and PstI)  scFv-C[J23101+B0034+OmpA+scFv-C+B030+E0040+J61048] Ar (EcoRI and PstI)   **Cultivation of**  GFP[J23101+B0034+E0040+J61048] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+E0040+J61048] Kr  scFv-C[R0010+B0034+OmpA+scFv-C]   **PCR of**  scFv-B[OmpA+scFv-B]  Anti-Her 2[OmpA+Herceptin-own]  GBP[FadL-GBP] Ar   **Transform of**  scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048]  scFv-B[J23101+B0034+OmpA+scFv-B +B0030+E0040+J61048]  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]   **Ligation of**  scFv-C[OmpA+scFv-C] Kr  scFv-B[OmpA+scFv-B] Kr  Anti-Her 2[OmpA+Herceptin-own] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Kr  scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048] Kr   **Plasmid purification of**  scFv-C[R0010+B0034+OmpA+scFv-C]  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+E0040+J61048]   **Digestion of**  scFv-C[R0010+B0034+OmpA+scFv-C] (EcoRI and PstI)  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+E0040+J61048] (EcoRI and PstI)  [pSB1K3] (EcoRI and PstI)  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own] (EcoRI and SpeI)  BFP[R0010+B0034+OmpA+K592100] (NcoI and PstI)   **Cultivation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own] Ar  scFv-B[OmpA+scFv-B] Kr  Anti-Her 2[OmpA+Herceptin-own] Kr  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]  OmpA[R0010+B0034+OmpA]   **Digestion of**  BFP[OmpA+K592100] (EcoRI and NcoI) | **26**   **Cultivation of**  [pSB1C3]  GBP[R0010+B0034+FadL-GBP+B0030+E0040] Kr   **Ligation of**  GBP[FadL-GBP+pSB1C3] Cr  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+E1010]  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+E0040]   **Electrophoresis of**  [pSB1K3] (EcoRI and PstI)  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own] (EcoRI and SpeI)  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+E0040]  scFv-C[R0010+B0034+OmpA+scFv-C]   **Digestion of**  GBP[FadL-GBP] (XbaI and PstI)   **Transform of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+E1010]  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own+B0030+K592100]   **Digestion of**  [J23101] (EcoRI and SpeI)  BFP[OmpA+K592100] (EcoRI and NcoI)   **Plasmid purification of**  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]  BFP[R0010+B0034+OmpA+K592100]  scFv-B[OmpA+scFv-B] Kr  Anti-Her 2[OmpA+Herceptin-own] Kr  [pSB1C3]   **Digestion of**  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] (EcoRI and PstI)  BFP[R0010+B0034+OmpA+K592100] (EcoRI and PstI)  scFv-B[OmpA+scFv-B] Kr(EcoRI and PstI I)  Anti-Her 2[OmpA+Herceptin-own] Kr (EcoRI and PstI)  [pSB1C3] | **27**  **Transform of**  GBP[FadL-LGBP] Cr   **Plasmid purification of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own]   **Ligation of**  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own] Cr  scFv-C[OmpA+scFv-C] Kr  scFv-B[OmpA+scFv-B] Kr  Anti-Her 2[OmpA+Herceptin-own] Kr  scFv-C[R0010+B0034+OmpA+scFv-C]  scFv-B [R0010+B0034+OmpA+scFv-B]  Anti-Her 2[R0010+B0034+OmpA+Herceptin-own]   **Digestion of**  GFP[B0034+E0040+J61048] (XbaI and PstI)  GBP[R0010+B0034+FadL-GBP] Ar (EcoRI and SpeI)  GFP[J23110+B0034+E0040+J61048] (EcoRI and PstI)  [pSB1C3] (EcoRI and PstI)   **Ligation of**  GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]  GFP[J23110+B0034+E0040+pSB6A1] | **28**  **PCR of**  GBP[FadL-GBP] Cr   **PCR of**  Anti-Her 2[J23110+B0034+Herceptin-own+B0030+E1010+J61048]  scFv-B[J23110+B0034+OmpA+scFv-B+B0030+K592100+J61048]  Anti-Her 2[J23110+B0034+Herceptin-own+B0030+K592100+J61048]  Anti-Her 2[OmpA+Herceptin-own]  scFv-C[J23110+B0034+OmpA+scFv-C]  Anti-Her 2[J23110+B0034+Herceptin-own]  scFv-C[OmpA+scFv-C].   **Transform of**  Anti-Her 2[J23110+B0034+Herceptin-own]  BFP[B0030+K592100+J61084]  BFP[B0030+K592100+J61084]  BFP[J23101+B0034+OmpA+K592100]   **Plasmid purification of**  scFv-B[OmpA+scFv-B] Kr  scFv-C[R0010+B0034+OmpA+scFv-C]   **Cultivation of**  Anti-Her 2[J23110+B0034+Herceptin-own+B0030+E0040+J61048]  Anti-Her 2[J23110+B0034+Herceptin-own+B0030] | **29** |
| **30**  **Plasmid purification of**  GBP[FadL-GBP]  Anti-Her 2[J23101+B0034+OmpA+Herceptin-own] | **31** |  |  |  |  |  |

[![](https://static.igem.org/mediawiki/2015/3/3c/%E7%AE%AD%E9%A0%AD1.png)

Back to Navigation](https://2015.igem.org/Team%3ANCTU_Formosa/Project)

[![](https://static.igem.org/mediawiki/2015/c/c2/%E7%AE%AD%E9%A0%AD2.png)

Go to Protocol](https://2015.igem.org/Team%3ANCTU_Formosa/Protocol)

* [Home](https://2015.igem.org/Team%3ANCTU_Formosa)
* + Project
  + [Description](https://2015.igem.org/Team%3ANCTU_Formosa/Description)
  + [Design](https://2015.igem.org/Team%3ANCTU_Formosa/Design)
  + [Results](https://2015.igem.org/Team%3ANCTU_Formosa/Results)
  + [Modeling](https://2015.igem.org/Team%3ANCTU_Formosa/Modeling)
  + [Safety](https://2015.igem.org/Team%3ANCTU_Formosa/Safety)
* + [Parts](https://2015.igem.org/Team%3ANCTU_Formosa/Parts)
  + [Basic Parts](https://2015.igem.org/Team%3ANCTU_Formosa/Basic_Part)
  + [Composite Parts](https://2015.igem.org/Team%3ANCTU_Formosa/Composite_Part)
* + Notebook
  + [Lab Safety](https://2015.igem.org/Team%3ANCTU_Formosa/LabSafety)
  + [Lab Note](https://2015.igem.org/Team%3ANCTU_Formosa/Notebook)
  + [Protocol](https://2015.igem.org/Team%3ANCTU_Formosa/Protocol)
* + Policy &
     Practices
  + [Human Practice](https://2015.igem.org/Team%3ANCTU_Formosa/Practices)
  + [Collaborations](https://2015.igem.org/Team%3ANCTU_Formosa/Collaborations)
* [Achievement](https://2015.igem.org/Team%3ANCTU_Formosa/Achievement)
* [Team](https://2015.igem.org/Team%3ANCTU_Formosa/Attributions)

![](https://static.igem.org/mediawiki/2015/5/54/Nctu_formosa_footer_team_logo.png)
![](https://static.igem.org/mediawiki/2015/b/b2/Nctu_formosa_logo123.png)

NCTU\_Formosa APOllO
[![](https://static.igem.org/mediawiki/2015/a/ab/NCTU_FORMOSA_Facebook_LINK.png)](https://www.facebook.com/pages/NCTU_Formosa-IGEM-team/267841893250331?fref=ts)
[National Chaio Tung University](http://www.nctu.edu.tw/)
Engineering Building 6 EF455, 1001 University Road, Hsinchu 300, Taiwan, ROC.