IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/30

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Team Allergy

  • Sequencing Results:

LTP ihpRNA parts that worked: 9c2, 11c1, 11c2

Bet 2 ihpRNA parts that worked: 25c1,c2, 28c1,(LTPS+PDK is actually Bet 2 hpRNA)

Ger ihpRNA parts that worked: 36c1,c2, 37c1,2, 38c1,c2

Team Flavor

  • Ligated StrepII tagged proteins into V24 backbone
  • Ligated cut J45004 into V0120 backbone

Ligations were plated and left @ 37°C O/N.

Team Fence

PCR of VP16 and Gal4DBD

1 2 3 4 5 6 7 8 9 10 11 12
KB+ Ladder Gal4 DBD 1 Gal4 DBD 2 Gal4 DBD 3 Gal4 DBD 4 VP16 1 VP16 2 VP16 3 VP16 4 KB+ Ladder

Image:PCR_VP16_Gal4DBD_7-30.jpg

PCR again for VP16

Using pfx polymerase

  • 1μL 1/10 dilution Hind.Fwd
  • 1μL 1/10 dilution VP16.Rev
  • 1μL DNTPs
  • 6μL enhancer buffer
  • .5μL MgSO4
  • .5μL PFX polymerase
  • 1μL 1/10 dilution VP16 #1
  • 5μL DH2O
  • 4μL amp(lification?) buffer

Running PCR on two reactions. There was not enough PFX for both to get .5μL, so only one of them is likely to actually amplify.

Second ACC + RxrHm digest gel

Image:sucesspcrkfkdigestacvtually.jpg

yes to ACC (left) no RxrHm



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