Virginia Commonwealth/31 July 2009

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Revision as of 15:45, 31 July 2009 by Trentay (Talk | contribs)

Contents

Tuesday 7 July 2009

Results

Maria and Afton

  • Overnight culture of I1352 grew, but no visible florescence
  • Transformation Results below
Name Plate Purpose Growth Observation
NEB 10 beta LB (+) Control shocked lawn
NEB 10 beta LB (+) Control lawn
NEB 10 beta AMP (-) Control no growth
NEB 10 beta CM (-) Control no growth
NEB 10 beta CM I1352 ligated to the backbone pSB4C5 no growth
NEB 10 beta AMP I1352 ligated to the backbone pSB4C5 hundreds of colonies; some expressing RFP
NEB 10 beta AMP I1352 ligated with itself hundreds of colonies; some expressing RFP
  • Interpretation of Transformation Results:
    • Shows the digestion/ligation process is not working
    • Because the I1352 grew successfully on an AMP plate after being 'ligated' with pSB4C5, shows that the original backbone is present
      • So it was not properly digested
    • No growth was visible on the CM plate with the ligated I1352 with the CM resistant backbone pSB4C5
      • If the digestion did not work properly, then the backbone could not be ligated with the I1352's promoter and RET
      • Also, all previous stocks of BioBricks with promoters (100-107,110,113,116,0011,0040) have successfully grown on CM resistance plates, meaning they have the correct backbone, but none express RFP.
    • Conclusion:
      • The enzymes are bad for digestion and possibly ligation
      • A new kit is being ordered
      • Plans need to be made to repeat the Digestion and Ligation process with all promoters when kit arrives

Trentay 15:38, 31 July 2009 (UTC)


Tasks

Afton and Maria


Wetlab

Afton and Maria