IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/29: Difference between revisions
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*The RxR-Hm did not show any evidence of the presence of an insert. | *The RxR-Hm did not show any evidence of the presence of an insert. | ||
*The RxR-Lc did show evidence of the presence of an insert, although it may be a bit larger than the expected size (600-700bp). Blurriness of the ladder makes it difficult to decipher. | *The RxR-Lc did show evidence of the presence of an insert, although it may be a bit larger than the expected size (600-700bp). Blurriness of the ladder makes it difficult to decipher. | ||
===Nanodrop of Gal4DBD Minipreps (from the beginning of the summer=== | |||
===PCR of VP16 and Gal4DBD (preparing for yeast vector insertion)=== | |||
Gal4DBD | |||
*2μL DNTPs | |||
*.5μL phusion polymerase | |||
*4μL 5x phusion buffer | |||
*1μL 1/10 dilution of BB.Gal4DBD.Rev | |||
*1μL 1/10 dilution of Bam.Gal4.Fwd | |||
*1μL 1/2 dilution Gal4DBD #3 (made 1/2 dilution such that 1μL will be <10 ng/μL | |||
*μL 10.5μLDH<sub>2</sub>O | |||
4 tubes of each, 20μL each tube | |||
==Team Flavor== | ==Team Flavor== |
Revision as of 14:03, 1 August 2010
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Team Allergy
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 FenceDigest of Wednesday's miniprepsSamples 1-9
insert sizes:
Samples 10-18
insert sizes
Samples 19-24
insert size
Summary
Nanodrop of Gal4DBD Minipreps (from the beginning of the summerPCR of VP16 and Gal4DBD (preparing for yeast vector insertion)Gal4DBD
4 tubes of each, 20μL each tube Team FlavorMinipreps
Gel Extraction
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