Team:Hawaii/Notebook/2008-07-14

From 2008.igem.org

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Things we did today

Wetlab work

PCR

Grace
  • 25 μl PCR reactions for GFP site directed mutagenesis and extraction of Biobrick pertinent sites from BBa_B0034

Made stocks

Grace
  • Made two 500mL batches of SOB
  • Made 20 LB + amp100 plates

Transformed DH5α with ligations of a Biobrick vector with pnir, slr2016-1, slr2016-2, pilA, or C0012

Grace
  • Transformed using 5 μl of 1:1 and 1:10 ligation dilutions
  • Transformed using 1 μl vector only (negative control)
  • Incubated on ice 10 min as opposed to the recommended 30 min. by iGEM

RE digested PCR products and plasmids

Grace
  • XbaI and PstI: GFP fusion brick and BBa_C0012
  • Added XbaI and PstI separately
  • Ran 25 μl of GFP and 20 μl of BBa_C0012 restriction digest reactions in gel
  • HindIII and BamHI: Biobrick segment and pRL1383a
  • Ran 25 μl of Biobrick segment (half of total rxn) and 20 μl of pRL1383a restriction digest reactions in gel
  • pRL1383a was not visbile under long wave UV. Need to rerun gel tomorrow with much more plasmid

Transformed DB3.1

Margaret
  • Transformed: pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-)nothing. The positive and negative control verify that the DB3.1 competent cells made on 7/11/08 can take up DNA. The results of this test can be seen here.

Discussion

Quote of the Day


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