iGEM 2017 — Team CCU_Taiwan

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# Team:CCU Taiwan/Notebook

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# [Notebook](https://2017.igem.org/Team%3ACCU_Taiwan)

* [Home](https://2017.igem.org/Team%3ACCU_Taiwan)
* Project
  + [Description](https://2017.igem.org/Team%3ACCU_Taiwan/Description)
  + [Biosensor](https://2017.igem.org/Team%3ACCU_Taiwan/Design)
    - [CSP detector](https://2017.igem.org/Team%3ACCU_Taiwan/Design#CSP)
    - [Lactate detector](https://2017.igem.org/Team%3ACCU_Taiwan/Design#Lactate)
  + [Test paper](https://2017.igem.org/Team%3ACCU_Taiwan/Test_paper)
  + [Experiments](https://2017.igem.org/Team%3ACCU_Taiwan/Experiments)
  + [Results](https://2017.igem.org/Team%3ACCU_Taiwan/Results)
  + [Demonstrate](https://2017.igem.org/Team%3ACCU_Taiwan/Demonstrate)
  + [Applied design](https://2017.igem.org/Team%3ACCU_Taiwan/Applied_Design)
  + [InterLab](https://2017.igem.org/Team%3ACCU_Taiwan/InterLab)
  + [Notebook](https://2017.igem.org/Team%3ACCU_Taiwan/Notebook)
* Hardware
  + [Hardware overview](https://2017.igem.org/Team%3ACCU_Taiwan/Hardware)
  + [Device design](https://2017.igem.org/Team%3ACCU_Taiwan/Device_design)
  + [Device detection](https://2017.igem.org/Team%3ACCU_Taiwan/Device_detection)
* Software
  + [IOT system](https://2017.igem.org/Team%3ACCU_Taiwan/IOT%20system)
  + [APP](https://2017.igem.org/Team%3ACCU_Taiwan/APP)
  + [Machine learning](https://2017.igem.org/Team%3ACCU_Taiwan/Machine_learning)
* [Modeling](https://2017.igem.org/Team%3ACCU_Taiwan/Model)
* Human practice
  + [Human practice overview](https://2017.igem.org/Team%3ACCU_Taiwan/Human_Practices)
  + [Silver HP](https://2017.igem.org/Team%3ACCU_Taiwan/HP/Silver)
  + [Integrate & Gold](https://2017.igem.org/Team%3ACCU_Taiwan/HP/Gold_Integrated)
  + [Public engagemant](https://2017.igem.org/Team%3ACCU_Taiwan/Engagement)
* [Entrepreneurship](https://2017.igem.org/Team%3ACCU_Taiwan/Entrepreneurship)
* [Safety](https://2017.igem.org/Team%3ACCU_Taiwan/Safety)
* Parts
  + [Parts](https://2017.igem.org/Team%3ACCU_Taiwan/Parts)
  + [Basic parts](https://2017.igem.org/Team%3ACCU_Taiwan/Basic_Part)
  + [Composite parts](https://2017.igem.org/Team%3ACCU_Taiwan/Composite_Part)
* Team
  + [Team members](https://2017.igem.org/Team%3ACCU_Taiwan/Members)
  + [Attributions](https://2017.igem.org/Team%3ACCU_Taiwan/Attributions)
  + [Collaborations](https://2017.igem.org/Team%3ACCU_Taiwan/Collaborations)
* [Medals](https://2017.igem.org/Team%3ACCU_Taiwan/Medals)

* [Week 01](#Week1)
* [Week 02](#Week2)
* [Week 03](#Week3)
* [Week 04](#Week4)
* [Week 05](#Week5)
* [Week 06](#Week6)
* [Week 07](#Week7)
* [Week 08](#Week8)
* [Week 09](#Week9)
* [Week 10](#Week10)
* [Week 11](#Week11)
* [Week 12](#Week12)
* [Week 13](#Week13)
* [Week 14](#Week14)
* [Week 15](#Week15)
* [Week 16](#Week16)

### Week 01 (2017/07/02~2017/07/08)

1. Lactate detection system

          Gene design of lactate detector

1. CSP detection system

          Gene design of CSP detector

### Week 02 (2017/07/09~2017/07/15)

1. Lactate detection system

          Gene design of lactate detector

1. CSP detection system

          Gene design of CSP detector
          Request 2 backbone plasmids of *Bacillus subtilis* from iGEM

| Part requesting #1 | BBa\_K823022 | pSBBs4S |
| --- | --- | --- |
| Part requesting #2 | BBa\_K823023 | pSBBs1C |

          Order chemical materials for preparing competent cell of *Bacillus subtilis*

### Week 03 (2017/07/16~2017/07/22)

1. Lactate detection system

          Gene design of lactate detector

1. CSP detection system

          Gene design of CSP detector

### Week 04 (2017/07/23~2017/07/29)

1. Lactate detection system

          Send orders to IDT for DNA synthesis

1. CSP detection system

          Gene design of CSP detector

### Week 05 (2017/07/30~2017/08/05)

1. Participate the 5th iGEM Asia-Pacific Conference at NCTU

### Week 06 (2017/08/06~2017/08/12)

1. Lactate detection system

1. CSP detection system

          Prepare competent cell of *Bacillus subtilis*
          - Amplify two backbone plasmids of *Bacillus subtilis*

1. InterLab Study

          Transform eight kinds of plasmids into *E. coli DH5α*

| Positive Control | BBa\_I20270 |
| --- | --- |
| Negative Control | BBa\_R0040 |
| Test Device 1 | BBa\_J364000 |
| Test Device 2 | BBa\_J364001 |
| Test Device 3 | BBa\_J364002 |
| Test Device 4 | BBa\_J364003 |
| Test Device 5 | BBa\_J364004 |
| Test Device 6 | BBa\_J364005 |

          Plate culture eight kinds of *E. coli DH5α*

### Week 07 (2017/08/13~2017/08/19)

1. Lactate detection system

          Amplify the IDT plasmid of lactate detection system

| IDT plasmid # 1 | LldR gene on IDT vector |
| --- | --- |
| IDT plasmid # 2 | GFP gene on IDT vector |
| IDT plasmid # 3 | MazEF gene on IDT vector |

1. CSP detection system

          Amplify the IDT plasmid of CSP detection system

| IDT plasmid # 1 | ComE gene on IDT vector |
| --- | --- |

### Week 08 (2017/08/20~2017/08/26)

1. Lactate detection system

          Restriction map analysis of IDT Plasmid

1. CSP detection system

          Restriction map analysis of *B. subtilis* backbone plasmid

### Week 09 (2017/08/27~2017/09/02)

1. Lactate detection system

          Function test of kill switch (mazEF)

1. CSP detection system

          Transformation test of pSBBs1C and pSBBs4S

### Week 10 (2017/09/03~2017/09/09)

1. Lactate detection system

          Amplify the backbone plasmid pSB1A2

1. CSP detection system

          Transformation test of pSBBs1C and pSBBs4S
          Amplify the IDT plasmid of CSP detection system

| IDT plasmid # 2 | ComD gene on IDT plasmid |
| --- | --- |
| IDT plasmid # 3 | GFP gene on IDT plasmid |

1. Test paper experiment

          Small scale culture with test paper
          Freeze dry the test paper

### Week 11 (2017/09/10~2017/09/16)

1. Lactate detection system

          Restriction map analysis of pSB1A2

1. CSP detection system

          Restriction map analysis of IDT plasmid

1. InterLab study

          Construct OD600 reference point
          Construct fluorescence standard curve

### Week 12 (2017/09/17~2017/09/23)

1. Lactate detection system

          pSB1A2\_GFP\_LldR assembly
          pSB1C3\_mazEF assembly

1. CSP detection system

          pSBBS1C\_comE assembly

1. InterLab study

          OD600 and fluorescence measurement of cell

### Week 13 (2017/09/24~2017/09/30)

1. Lactate detection system

          Restriction map analysis of recombination DNA

1. CSP detection system

          pSBBs1C\_comD assembly

1. InterLab study

          E-mail InterLab data to iGEM
          Fill out three online form of InterLab

1. Test paper experiment

          Culture the bacteria with 600 mL LB in Erlenmeyer flask which contains test paper
          Freeze dry the test paper

### Week 14 (2017/10/01~2017/10/07)

1. Lactate detection system

          Function test of lactate detector
          Function test of kill switch

1. CSP detection system

          pSBBs1C\_comE\_comD assembly
          pSBBs4S\_GFP assembly

1. Test paper experiment

          Culture the bacteria with 600 mL LB in Erlenmeyer flask which contains test paper
          Freeze dry the liquid culture

### Week 15 (2017/10/08~2017/10/14)

1. Lactate detection system

          Double plasmid transformation test
          Function test of lactate detector
          Function test of kill switch

1. CSP detection system

          Check the size of assembly plasmid
          Transform the recombination plasmid to *B. subtilis*

1. Test paper experiment

          Revive the freeze-dried bacteria

### Week 16 (2017/10/15~2017/10/21)

1. Lactate detection system

          Bacteria which contain two plasmid

1. CSP detection system

          Prepare competent cell of *B. subtilis*
          Transform the recombination plasmid to *B. subtilis*

1. Test paper experiment

          Freeze dry the liquid culture, and then revive it

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