IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/29: Difference between revisions

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ACC synthase: 300 to 350
ACC synthase: 300 to 350
Terminator:  
Terminator: unknown
Act2LacI: ~1200
Act2LacI: ~1200



Revision as of 11:22, 29 July 2010

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Team Allergy

  • Ran a gel for our second attempt at amiRNA stiching (stage 1): . It seems that the backbone is still present at high concentrations after PCR purification. Maybe this is why last time we had a high incidence of background? This time we're gel purifying.
  • Dignostic Digest

Gel: ladder, 9, 9, 11, 11, 25, 25, 28, 28, ladder, 36, 36, 37, 37, 38, 38, ladder, LTPS+ PDK, LTPS+PDK, Bet2S+PDK, Bet2S+PDK, GerS+PDK, GerS+PDK, GerS+PDK, GerS+PDK

Expected band lengths:

Complete Parts

9,11 (LTPS+Pal+LTPA): 840bp and 3200 bp

25,28 (Bet2S+Pal+Bet2A): 840bp and 3200 bp

36,37,38 (GerS+Pal+GerA): 840bp and 3200 bp

Sense + Intron Parts

LTPS+ PDK: 1200 and 3200 bp

Bet2S+ PDK: 1200 and 3200 bp

GerS+PDK: 1200 and 3200 bp

(9 c1, 25 c2, 36 c1 have a 540 and 3200 band as opposed to a 840 and 3200 band--prbly b/c sense+intron is there but antisense was not ligated in). All intron+sense parts except for LTPS+PDK appear to have 300 bands as well indicating that PDK was not ligated in.

(a small mix up with two of the lids was made and its possible that LTPS+PDK (w/ an 840 and 3200 band is actually the complete Bet2 part so we're sending this in for sequening anyway).

Team Fence

Digest of Wednesday's minipreps

  • Lane 1: ladder
  • Lane 2: ladder
  • Lane 3: ACC synthase 1
  • Lane 4: ACC synthase 2
  • Lane 5: Acc synthase 3
  • Lane 6: Terminator 1
  • Lane 7: Terminator 2
  • Lane 8: Terminator 3
  • Lane 9: Act2LacI 1
  • Lane 10: Act2LacI 2
  • Lane 11: Act2LacI 3
  • Lane 12: ladder

insert sizes:

ACC synthase: 300 to 350 Terminator: unknown Act2LacI: ~1200

Team Flavor

Minipreps

  • Miniprep of Mira/Brazz+StrepII+STOP in V0120

Gel Extraction

  • Gel extraction of J45017: upcoming