Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/17
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Revision as of 18:24, 22 October 2010
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E.coli Fiber Project Notebook
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2010/8/17 Tuesday (watachin)
Experiment:Electrophoreses of PCR productions
Member
NEX and watachin
Materials
- pSB1A3(25ng/μl) 22μl
- 10*Loading buffer 2.2μl
- DNA Marker 5μl
- 1*TAE buffer
- 1% agarose gel
Procedure
- Set agarose gel and add TAE buffer in electrophoresis tank.
- Mix Loading Buffer and pSB1C3 ,then put them in well (marker sets another well).
- Load DNA at 100V for two thirds of total volume (about 15 minutes).
- Image the consequence of electrophoresis.
Result
failure
Plasmid concentration was too low.
Experiment:Transformation of pSB1A3
Member
NEX and watachin
Material
- pSB1A3 1μl
- competent cell DH5α 50μl
- LB + amp plate
Procedure
- mix pSB1A3 and DH5α
- on ice (30min)
- heat shock 42℃ 45sec
- on ice (2min)
- inoculate this onto plate
- incubate cells at 37℃