Team:Stockholm/5 October 2010
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==Andreas== | ==Andreas== | ||
+ | ===Transfer of pEX.nCPP⋅SOD⋅His to BL21=== | ||
+ | ====Glycerol stocks==== | ||
+ | *BL21 pEX.nLMWP⋅SOD⋅His | ||
+ | *BL21 pEX.nTra10⋅SOD⋅His | ||
+ | |||
+ | ===Assembly of His⋅SOD⋅cCPP constructs=== | ||
+ | ====Colony PCR==== | ||
+ | #pSB1C3.His⋅SOD⋅cTra10 A-D | ||
+ | #pSB1C3.His⋅SOD⋅cTAT A-D | ||
+ | #pSB1C3.His⋅SOD⋅cLMWP A-D | ||
+ | |||
+ | Standard colony PCR settings | ||
+ | *Elongation time: 1:30 | ||
+ | |||
+ | ====Gel verification==== | ||
+ | 1 % agarose, 120 V | ||
+ | |||
+ | '''Expected bands''' | ||
+ | #884 bp | ||
+ | #854 bp | ||
+ | #863 bp | ||
+ | |||
+ | '''Results'''<br /> | ||
+ | Varying band sized, with the following clones seeming correct: | ||
+ | #A | ||
+ | #D | ||
+ | #B & C |
Revision as of 12:27, 5 October 2010
Contents |
Andreas
Transfer of pEX.nCPP⋅SOD⋅His to BL21
Glycerol stocks
- BL21 pEX.nLMWP⋅SOD⋅His
- BL21 pEX.nTra10⋅SOD⋅His
Assembly of His⋅SOD⋅cCPP constructs
Colony PCR
- pSB1C3.His⋅SOD⋅cTra10 A-D
- pSB1C3.His⋅SOD⋅cTAT A-D
- pSB1C3.His⋅SOD⋅cLMWP A-D
Standard colony PCR settings
- Elongation time: 1:30
Gel verification
1 % agarose, 120 V
Expected bands
- 884 bp
- 854 bp
- 863 bp
Results
Varying band sized, with the following clones seeming correct:
- A
- D
- B & C