Team:Tokyo Metropolitan/Notebook/Fiber/2010/10/19

E.coli Fiber Project Notebook

2010/10/19 Tuesday (Naoto)

member naoto and watachin

Experiment:Colony PCR

material

• colony of E.coli
  • bcsB:1~96
  • bcsC:97~196

other materials were same as protocol3

procedure

see protocol3

Experiment:Electrophoresis

material

• PCR production (Colony PCR)

you can see other materials at protocol4

procedure

refer to protocol4

result

We couldn't see bands of bcsB and bcsC...

Experiment:Sequence

material

• bcsD in pSB1C3(No.89~96)
• Big Dye
• primer
• DW
• ethanol
• EDTA

procedure

1. mix materials(DNA<50ng)
2. PCR
3. Add EDTA and Ethanol and put at room temperature　(15min)
4. centrifuge (8000rpm,30min)
5. throw away supernatant
6. add ethanol again
7. centrifuge (8000rpm,15min)
8. put at refrigerator
9. throw away supernatant
10. add Hi-Di solution
11. transfer these sample to plate for sequence
12. read sequence

result

Failure to read sequence