Team:Stockholm/22 September 2010

From 2010.igem.org

(Difference between revisions)
(Andreas)
(Andreas)
Line 2: Line 2:
==Andreas==
==Andreas==
===Gel verification===
===Gel verification===
 +
[[image:ColPCR_newassemblies22sep.png‎|200px|thumb|right|'''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.'''<br />4 &mu;l &lambda;; 4 &mu;l sample.<br />&lambda; = O'GeneRuler 1 kb DNA ladder.]]
''Of 21/9 colony PCR samples''
''Of 21/9 colony PCR samples''
Line 7: Line 8:
'''Expected bands'''
'''Expected bands'''
-
*pEX.N-TAT&sdot;SOD&sdot;His (pEX.nTSH): 735 bp
 
*pSB1A2.RBS&sdot;yCCS (pA.RBSy): 1018 bp
*pSB1A2.RBS&sdot;yCCS (pA.RBSy): 1018 bp
 +
*pEX.N-TAT&sdot;SOD&sdot;His (pEX.nTSH): 735 bp
*pSB1C3.N-LMWP&sdot;SOD&sdot;His (pC.nLSH): 857 bp
*pSB1C3.N-LMWP&sdot;SOD&sdot;His (pC.nLSH): 857 bp
*pSB1K3.N-LMWP&sdot;SOD&sdot;His (pK.nLSH): 857 bp
*pSB1K3.N-LMWP&sdot;SOD&sdot;His (pK.nLSH): 857 bp
Line 14: Line 15:
'''Results'''
'''Results'''
 +
*pSB1A2.RBS&sdot;yCCS: Seemingly correct-sized bands for '''clones 2, 3 and 4'''.
 +
*pEX.N-TAT&sdot;SOD&sdot;His: Seemingly correct-sized bands for '''clones 3 & 4'''.
 +
*pSB1C3.N-LMWP&sdot;SOD&sdot;His: Seemingly correct-sized bands for '''clones 1 & 4'''.
 +
*pSB1K3.N-LMWP&sdot;SOD&sdot;His: Seemingly correct-sized band for '''clone 1'''.
 +
 +
===ON cultures===
 +
#5 ml LB + appr. antibiotic (100 Amp, 25 Cm or 50 Km); 37 &deg;C, 225 rpm
 +
#*pSB1A2.RBS&sdot;yCCS: clones 2, 3 and 4
 +
#*pEX.N-TAT&sdot;SOD&sdot;His: clones 3 and 4
 +
#*pSB1C3.N-LMWP&sdot;SOD&sdot;His: clones 1 and 4
 +
#*pSB1K3.N-LMWP&sdot;SOD&sdot;His: clone 1

Revision as of 15:10, 22 September 2010


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 2, 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