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Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/01
From 2010.igem.org
E.coli Fiber Project Notebook
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Contents |
2010/9/1 Wednesday(Naoto)
Experiment:digestion of bcsA,B,C,D(A.xylinum)
member
naoto
material
- bcsA,B,C,D(from A.xylinum)
- 10×Mbuffer
- BSA
- XbaI
- SpeI
procedure
- add "material" to PCR tubes(show below)
- incubation at (37℃,7h)
| bcsA | bcsB | bcsC | bcsD | ||
|---|---|---|---|---|---|
| solution of bcs(μl) | 20 | 20 | 20 | 20 | |
| 10×Mbuffer(μl) | 2 | 2 | 2 | 2 | |
| BSA(μl) | 2 | 2 | |||
| XbaI(μl) | 0.8 | 0.8 | |||
| SpeI(ul) | 0.8 | 0.8 |
Experiment:subculture ofA.xylinum
member
naoto
material
A.xylinum JCM7664
procedure
transfer A.xylinum JCM7664 to new culture(OWW and JCM Broth 8/25 made)
Experiment:subculture of E.coli
member
naoto
material
E.coli K12
procedure
pick up a culture of E.coli K12 and streak it to new culture(LB plate)
Experiment:direct PCR
member
naoto
material
- E.coli K12
- 2×PCR buffer 25μl×2
- 2mM dNTP 10μl×2
- 10μM primer(sense)bcsA,B 2.5μl each
- 10μM primer(antisense)bcsA,B 2.5μl each
- milli-Q water 9μl×2
- KOD FX 0.5μl×2
procedure
follow protocol3 direct PCR
