Team:Stockholm/5 October 2010

From 2010.igem.org

(Difference between revisions)
(Andreas)
(Andreas)
Line 16: Line 16:
====Gel verification====
====Gel verification====
 +
[[image:ColPCR_HS*cCPP_5oct.png|200px|thumb|right|'''Colony PCR gel verification of His&sdot;SOD&sdot;cCPP constructs in pSB1C3.'''<br />4 &mu;l &lambda;; 5 &mu;l sample.<br />&lambda; = O'GeneRuler 1 kb DNA ladder.]]
1 % agarose, 120 V
1 % agarose, 120 V

Revision as of 12:37, 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

  1. pSB1C3.His⋅SOD⋅cTra10 A-D
  2. pSB1C3.His⋅SOD⋅cTAT A-D
  3. pSB1C3.His⋅SOD⋅cLMWP A-D

Standard colony PCR settings

  • Elongation time: 1:30

Gel verification

Colony PCR gel verification of His⋅SOD⋅cCPP constructs in pSB1C3.
4 μl λ; 5 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.

1 % agarose, 120 V

Expected bands

  1. 884 bp
  2. 854 bp
  3. 863 bp

Results
Varying band sized, with the following clones seeming correct:

  1. A
  2. D
  3. B & C