Team:Stockholm/2 October 2010
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====Gel verification==== | ====Gel verification==== | ||
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+ | [[image:ColPCR_pEX.CPP_SYoperon_2oct.png|200px|thumb|right|'''Colony PCR gel verification of BL21 pEX.nCPP*SOD*His constructs and Top10 pEX.nCPP*SOD*His.RBS.yCCS operon constructs.'''<br />4 μl λ; 4 μl sample.<br />λ = O'GeneRuler 1 kb DNA ladder.]] | ||
0.8 % agarose, 100 V | 0.8 % agarose, 100 V | ||
Revision as of 15:41, 2 October 2010
Contents |
Andreas
Transfer of nCPP⋅SOD⋅His.RBS.yCCS operon to pEX
Continued from 30/8 transformations
Colony PCR
- BL21 pEX.nLMWP⋅SOD⋅His: A & B
- BL21 pEX.nTra10⋅SOD⋅His: A & B
- pEX.nTAT⋅SOD⋅His.RBS.yCCS 2: A & B
- pEX.nTAT⋅SOD⋅His.RBS.yCCS 3: A & B
- pEX.nTra10⋅SOD⋅His.RBS.yCCS 1: A & B
- pEX.nTra10⋅SOD⋅His.RBS.yCCS 2: A & B
- pEX.nLMWP⋅SOD⋅His.RBS.yCCS 2: A & B
- pEX.nLMWP⋅SOD⋅His.RBS.yCCS 3: A & B
Standard colony PCR settings.
- Elongation time: 2:00
Gel verification
0.8 % agarose, 100 V
Expected bands:
- 744 bp
- 765 bp
- 1523 bp
- 1523 bp
- 1553 bp
- 1553 bp
- 1532 bp
- 1532 bp
Results
Ran gel a little too long too see correct clones of constructs 1 and 2. For the other it looks like many clones have double inserts, probably a result of too high insert:vector ratio during ligation.
Confirmed clones:
- -
- -
- A
- B
- -
- -
- B
- B
These will be saved for later.