iGEM 2014 — Team OUC-China

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# Team:OUC-China/Notebook

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  + [Overview](https://2014.igem.org/Team%3AOUC-China/Project)
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* NOTEBOOK
  + [Lab note](https://2014.igem.org/Team%3AOUC-China/Notebook)
  + [Modeling note](https://2014.igem.org/Team%3AOUC-China/Notebook_Modeling_Note)
  + [Protocols](https://2014.igem.org/Team%3AOUC-China/Notebook_Protocols)

# Lab Note

WEEK 1

We acquired foundational knowledge as preparation for our project.Under the guidance of teachers and fellow students, we studied molecular biology and synthetic biology.

WEEK 2

Then we learned molecular experiments, included bacterination, plasmid miniprep, agarose electrophoresis, enzyme digestion, gel extraction, ligation, transformation, bacterial coating, PCR and so on.

WEEK 3

We acquired plasmid RP4 fromother lab,professor QIU Zhigang provided us HB101 strain which contains plasmid RP4. After getting HB101 strain, we tested the plasmid in several ways. In the end, the test results show the plasmid is exactly what we want. Firstly, through resistance screening experiment to determine if the types of antibiotics genes carried by the plasmid are consistent with literature. Secondly, getting the vital sequence from the plasmid by means of PCR to confirm whether the plasmid is RP4 or not.Thirdly, by doing conjugation experiment to test if the plasmid could conjugate with other strains.

We built a plasmid which used pet32a as backbone and the gene sequence of TAT-H4, and then we transformed it into BL21 (DE3).

Experiment:

a) Transform：transform our plasmid into BL21(DE3) and culture it on LB solid medium.

b) Plasmid miniprep: plasmid extraction from BL21 (DE3).

c) Agarose electrophoresis: detect length of the extracted plasmid, it’s about 6000bp so the it confirms the correctness of the result.

d) Culture: HB101 strain wasscreened by ampicillin and kanamycinrespectivelyin LB broth. And both bottles of

broth were turbid.

e) plasmid miniprep: plasmid extraction from HB101.

f) agaroseelectrophoresis: detect length of the extracted plasmid, and the result is right.

![](https://static.igem.org/mediawiki/2014/f/ff/OUC-China-1-RP4.png)

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