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Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/01
From 2010.igem.org
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==2010/9/1 Wednesday(Naoto)== | ==2010/9/1 Wednesday(Naoto)== | ||
| - | ===Experiment: | + | ===Experiment:Digestion of bcsA,B,C and D=== |
| - | ''' | + | :'''Member''' |
| - | naoto | + | :naoto |
| - | ''' | + | :'''Material''' |
| - | *bcsA,B,C,D | + | :*bcsA,B,C,D |
| - | *10×Mbuffer | + | :*10×Mbuffer |
| - | *BSA | + | :*BSA |
| - | *XbaI | + | :*XbaI |
| - | *SpeI | + | :*SpeI |
| - | ''' | + | :'''Procedure''' |
#add "material" to PCR tubes(show below) | #add "material" to PCR tubes(show below) | ||
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|} | |} | ||
| + | ===Preparation:Subculture of''A.xylinum''=== | ||
| + | :'''Member''' | ||
| - | + | :naoto | |
| - | + | ||
| - | + | :'''Material''' | |
| - | '' | + | :''A.xylinum'' JCM7664 |
| - | '' | + | :'''Procedure''' |
| - | '' | + | :transfer ''A.xylinum'' JCM7664 to new culture(OWW and Broth 8/25 made) |
| - | + | ===Experiment:PCR=== | |
| + | :'''Member''' | ||
| - | + | :naoto | |
| - | + | ||
| - | + | :'''Material''' | |
| - | + | ||
| - | ''' | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
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| + | :* ''E.coli'' K12 strain | ||
| + | :* 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 <html><a href="https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Fiber/Protocol#.E3.80.80PCR">protocol3</a></html> | ||
<br/> | <br/> | ||
Revision as of 22:01, 26 October 2010
E.coli Fiber Project Notebook
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2010/9/1 Wednesday(Naoto)
Experiment:Digestion of bcsA,B,C and D
- Member
- naoto
- Material
- bcsA,B,C,D
- 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 |
Preparation:Subculture ofA.xylinum
- Member
- naoto
- Material
- A.xylinum JCM7664
- Procedure
- transfer A.xylinum JCM7664 to new culture(OWW and Broth 8/25 made)
Experiment:PCR
- Member
- naoto
- Material
- E.coli K12 strain
- 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
