Team:Tokyo Metropolitan/Notebook/Fiber/2010/08/17

E.coli Fiber Project Notebook

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

1. Set agarose gel and add TAE buffer in electrophoresis tank.
2. Mix Loading Buffer and pSB1C3 ,then put them in well (marker sets another well).
3. Load DNA at 100V for two thirds of total volume (about 15 minutes).
4. 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

1. mix pSB1A3 and DH5α
2. on ice (30min)
3. heat shock 42℃ 45sec
4. on ice (2min)
5. inoculate this onto plate
6. incubate cells at 37℃