"
Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/15
From 2010.igem.org
E.coli Fiber Project Notebook
|
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Contents |
2010/9/15 Wednesday (Naoto)
Experiment:Electrophoresis
member
naoto and watachin
material
- PCR production (pSB1C3)
If you want to know other materials, see protocol4
procedure
see protocol4
result
bands could be seen slightly
Experiment:subculture of A.xylinum
member
naoto and watachin
material
see protocol1 A.xylinum(subcultured at 8/24 and 9/1)
procedure
see protocol1 we made 3 OWW plates and 3 tubes of broth
Experiment:Extraction of DNA from agarose gel with QIAGEN
member
naoto and watachin
material
- pSB1C3 in agarose gel
If you want to know other materials, see protocol4
procedure
see protocol5
Experiment:PCR
member
naoto and watachin
material
- pSB1C3,bcsB(A.xylinum)
- a culture of E.coli
If you want to know other materials, see protocol3(use KOD pol)
procedure
- add materials to PCR tubes
- PCR reaction (see below)
- nitialization :95°C 3min
- denaturation:96°C 1min
- annealing :51°C 1min
- elongation:72°C 3min
(30cycles from 96°C 1min to 72°C 3min)
- reaction stop :10°C ∞
Experiment:Electrophoresis
member
naoto and watachin
material
- PCR production (see above PCR)
If you want to know other materials, see protocol4
procedure
see protocol4
result
a band of pSB1C3 could be seen slightly bands of bcsB(A.xylinum),bcsC(E.coli) appeared clearly
Experiment:Digestion
member naoto and watachin
material
- pSB1C3,bcsA(A.xylinum) and bcsC(E.coli)
- 10×Mbuffer
- BSA
- XbaI
- SpeI
| pBS1C3 | bcsA | bcsC | ||
|---|---|---|---|---|
| solution of bcs(μl) | 50 | 50 | 50 | |
| 10×Mbuffer(μl) | 5 | 5 | 5 | |
| BSA(μl) | 5 | 5 | 5 | |
| XbaI(μl) | 1 | 1 | 1 | |
| SpeI(ul) | 1 | 1 | 1 |
procedure
- add "material" to PCR tubes(show below)
- incubation at 37℃,14h
Experiment:Extraction of DNA from agarose gel
member
naoto and watachin
material
- pSB1C3,bcsB(A.xylinum) and bcsC(E.coli) in agarose gel
If you want to know other materials, see the protocol5
procedure
see If you want to know other materials, see the protocol5
