IGEM:IMPERIAL/2006/LabCalendar/2006-9-11

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TO DO:
*T9002 and J37016 testing
*Need to make more frozen stocks of J37016 ?
*CHECK & possibly repeat ligation for J37024
*Get PCR fusion from Kirsten & test CRE/Lox part
*Ask Kiron about our protein gel
*Sequence 5M, J37022new, (Dr. Fray's aiiA (primers!))

Contents

Frozen Stocks of J37016

  • Made more

Minprep results of J37024

  • Miniprep showed that the ligation has failed. However, we cannot explain the bands that appear on the gel...they don't seem to have come from the parental parts either...
  • A control miniprep was run as a check (should get results tomorrow). Minipreps from many more colonies on the plate should be done on Wednesday (couldn't culture today as we lost access to the lab).

Re-Ligation of J37024

  • The maxiprep digest showed strange results (four bands resulted from each digest). Dr. Mann and Kirsten suggested it may be due to a high concentration of DNA. We couldn't differentiate between some of the bands and so discontinued our ligations.
  • We used Dr. Mann's ligation instead (he had offered to do it in parallel). We electroporated half of the ligation today and left the other half to ligate overnight (giving us two chances...just in case...)

Protein Gel

  • Western blot showed no results at all...but this is inconclusive since the enzyme only had a 50-50 chance of being expressed in Rosetta.

T9002 + J37016 Testing

  • Carried out by Vincent.

Sequencing

  • To be done on Tuesday.

Testing Cre/Lox

  • To be done on Tuesday.
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