Team:Stockholm/22 September 2010

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(→Andreas)
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===ON cultures===
===ON cultures===
#5 ml LB + appr. antibiotic (100 Amp, 25 Cm or 50 Km); 37 °C, 225 rpm
#5 ml LB + appr. antibiotic (100 Amp, 25 Cm or 50 Km); 37 °C, 225 rpm
-
#*pSB1A2.RBS⋅yCCS: clones 2, 3 and 4
+
#*pSB1A2.RBS⋅yCCS: clones 3 and 4
#*pEX.N-TAT⋅SOD⋅His: clones 3 and 4
#*pEX.N-TAT⋅SOD⋅His: clones 3 and 4
#*pSB1C3.N-LMWP⋅SOD⋅His: clones 1 and 4
#*pSB1C3.N-LMWP⋅SOD⋅His: clones 1 and 4
#*pSB1K3.N-LMWP⋅SOD⋅His: clone 1
#*pSB1K3.N-LMWP⋅SOD⋅His: clone 1
 +
 +
::'''***Note to self***'''<br />
 +
::''It might be possible that pSB1A2.RBS&sdot;yCCS clones have been mixed up with pEX.N-TAT&sdot;SOD&sdot;His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes.

Revision as of 15:46, 22 September 2010

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Andreas

Gel verification

Colony PCR gel verification of new assemblies: pSB1A2.RBS.yCCS, pEX.N-TAT*SOD*His, pSB1C3.N-LMWP*SOD*His & pSB1K3.N-LMWP*SOD*His.
4 μl λ; 4 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.

Of 21/9 colony PCR samples

1 % agarose, 120 V

Expected bands

  • pSB1A2.RBS⋅yCCS (pA.RBSy): 1018 bp
  • pEX.N-TAT⋅SOD⋅His (pEX.nTSH): 735 bp
  • pSB1C3.N-LMWP⋅SOD⋅His (pC.nLSH): 857 bp
  • pSB1K3.N-LMWP⋅SOD⋅His (pK.nLSH): 857 bp
  • pSB1C3.SOD⋅His (PC): 815 bp

Results

  • pSB1A2.RBS⋅yCCS: Seemingly correct-sized bands for clones 2, 3 and 4.
  • pEX.N-TAT⋅SOD⋅His: Seemingly correct-sized bands for clones 3 & 4.
  • pSB1C3.N-LMWP⋅SOD⋅His: Seemingly correct-sized bands for clones 1 & 4.
  • pSB1K3.N-LMWP⋅SOD⋅His: Seemingly correct-sized band for clone 1.

ON cultures

  1. 5 ml LB + appr. antibiotic (100 Amp, 25 Cm or 50 Km); 37 °C, 225 rpm
    • pSB1A2.RBS⋅yCCS: clones 3 and 4
    • pEX.N-TAT⋅SOD⋅His: clones 3 and 4
    • pSB1C3.N-LMWP⋅SOD⋅His: clones 1 and 4
    • pSB1K3.N-LMWP⋅SOD⋅His: clone 1
***Note to self***
It might be possible that pSB1A2.RBS⋅yCCS clones have been mixed up with pEX.N-TAT⋅SOD⋅His. After plasmid prep, this should be tested by digesting the samples and analyze digestion sizes.
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