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Team:Stockholm/2 October 2010
From 2010.igem.org
(Difference between revisions)
(→Gel verification) |
(→Transfer of nCPP⋅SOD⋅His.RBS.yCCS operon to pEX) |
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''Continued from 30/8 transformations'' | ''Continued from 30/8 transformations'' | ||
====Colony PCR==== | ====Colony PCR==== | ||
| - | #<u>BL21</u> pEX.nLMWP⋅SOD⋅His: A & B | + | #<u>BL21</u> pEX.nLMWP⋅SOD⋅His: A & B † |
| - | #<u>BL21</u> pEX.nTra10⋅SOD⋅His: A & B | + | #<u>BL21</u> pEX.nTra10⋅SOD⋅His: A & B † |
#pEX.nTAT⋅SOD⋅His.RBS.yCCS 2: A & B | #pEX.nTAT⋅SOD⋅His.RBS.yCCS 2: A & B | ||
#pEX.nTAT⋅SOD⋅His.RBS.yCCS 3: A & B | #pEX.nTAT⋅SOD⋅His.RBS.yCCS 3: A & B | ||
| Line 11: | Line 11: | ||
#pEX.nLMWP⋅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 | #pEX.nLMWP⋅SOD⋅His.RBS.yCCS 3: A & B | ||
| + | |||
| + | † ''From "Transformation of BL21, 30/8" | ||
Standard colony PCR settings. | Standard colony PCR settings. | ||
Revision as of 14:04, 4 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
† From "Transformation of BL21, 30/8"
Standard colony PCR settings.
- Elongation time: 2:00
Gel verification
4 μl λ; 4 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.
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.
