Team:Northwestern/Notebook/Week9

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Revision as of 21:43, 15 August 2011 by Rafay (Talk | contribs)

RETURN TO IGEM 2010



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Day 39 - Monday, August 8th 2011

  • Miniprepped overnight cultures from the attempted Oligo based ligations last week.
  • Digested the minipreps and ran them on a gel to look for the correct sized bands.
  • Prepared samples for sequencing
  • Made new Tet Plates, Chlor Plates and SOB Broth
  • PCRed our strep tag primer onto our CP->LasR and CP-> RhlR constructs


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Day 40 - Tuesday, August 9th 2011

  • Made a new batch of competent cells. We tried to get them in the -80 freezer extremely quickly to improve the efficiency.
  • We digested our strep tag PCR products with DpnI and then added ligase so the blunt ends would come back together. We ligated one batch for one hour and another overnight.
  • We transformed the overnight ligations
  • We transformed to test the competency of our new competent cells
  • Sent in samples for seqencing
  • Designed our T-shirts!
  • Rafay gave a presentation on the math model


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Day 41 - Wednesday, August 10th 2011

  • Our competent cell test had only 21 colonies, indicating a very low efficiency =(
  • Our transformations of the strep tag ligations also failed, although we can still try transforming the overnight ligations.
  • Hopefully we’ll get sequencing results soon....
  • Miniprepped the overnights from our glycerol stocks and of our terminators
  • Transformed the strep tag overnight ligations from yesterday


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Day 42 - Thursday, August 11th 2011

  • The overnight ligations of the strep tag constructs produced four colonies! These were overnighted for miniprep tomorrow.
  • Ran another digest of the CP -> RBS -> LasR/RhlR -> Strep tag constructs, this time after phosphorylating the ends of the DNA. This should help improve ligation efficiency.
  • Ligated those digestions overnight
  • Ran our strep tag PCR products on a gel to try and confirm that the initial amplification was successful.
  • Prepared overnights of RBS+Part ligations from glycerol stocks and transformation plates. We will ligate our genomic promoters to these tomorrow.
  • Started 3 test ligations with Genomic Promoter -> RBS -> Part from already existing stocks.
  • Made a lot of progressing designing our human practices project
  • The team and sponsorship pages on the wiki are nearing completion.


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Day 43 - Friday, August 12th 2011

  • Transformed the strep tag ligations
  • Miniprepped overnights of RBS34+Part Ligations
  • We got our sequencing results back! Most of the samples look very good and closely match what we expected.
  • Made some more progress on the website.