IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/08/03: Difference between revisions
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==Team Fence== | ==Team Fence== | ||
VP16 PCR from last night | |||
[[Image: 83gelpcrvp16.jpg||350px]] | |||
===Yesterday's transformations=== | |||
[[Image:Gal4-EcR_lig_8-3.jpg||350px]] | |||
[[Image:EcR_neg_control_8-3.jpg||350px]] | |||
[[Image:LTP_lig-1_8-3.jpg||350px]] | |||
[[Image:LTP_lig-2_8-3.jpg||350px]] | |||
[[Image:B11_neg_cont_lig_8-3.jpg||350px]] | |||
[[Image:Barn-strep_lig_8-3.jpg||350px]] | |||
[[Image:B15_cont_lig_8-3.jpg|350px]] | |||
===Glycerol Stocks=== | |||
from 7-27-10 | |||
keeping Act2lac0pt 1, ACC synthase 1, EcR 2, RXRLc 2 | |||
added .5mL 80% glycerol to .5mL overnight cell culture of selected transformation, vortexed thoroughly, and placed in the -80°C freezer in team fence box. | |||
===Annealing 35s min promoter=== | |||
*3μL 100μM 35sminpromt.Rev | |||
*3μL 100μM 35sminpromt.Fwd | |||
*3μL .5M NaCl | |||
*3μL 10x PNK buffer | |||
*18μL DH<sub>2</sub>O | |||
placed eppendorf in boiling water for 2 mins, then removed water from heat and allowed water and eppendorf to slowly cool together. | |||
==Team Flavor== | |||
* Colonies were obtained from all ligations; 5mL cultures were started, left ~5hrs @ 37°C | |||
* Glycerol stocks were made (666μL glycerol: 333μL cells; 50% glycerol final) | |||
* Miniprepped per Qiagen protocol | |||
ODs | |||
004-1: 95.5 ng/μL | |||
004-2: 124.5 ng/μL | |||
017-1: 109.7 ng/μL | |||
017-2: 77.5 ng/μL | |||
Miraculin N+Strep+Stop: 61.5 ng/μL | |||
Miraculin C+Strep+Stop: 78.5 ng/μL | |||
Brazzein N+Strep+Stop: 83.4 ng/μL | |||
Brazzein C+Strep+Stop: 99.4 ng/μL | |||
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Revision as of 14:17, 7 September 2010
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Team Allergy
[[Image: |240px|PCR gel]] ladder, Team FenceYesterday's transformationsGlycerol Stocksfrom 7-27-10 keeping Act2lac0pt 1, ACC synthase 1, EcR 2, RXRLc 2 added .5mL 80% glycerol to .5mL overnight cell culture of selected transformation, vortexed thoroughly, and placed in the -80°C freezer in team fence box. Annealing 35s min promoter
placed eppendorf in boiling water for 2 mins, then removed water from heat and allowed water and eppendorf to slowly cool together. Team Flavor
ODs 004-1: 95.5 ng/μL 004-2: 124.5 ng/μL 017-1: 109.7 ng/μL 017-2: 77.5 ng/μL Miraculin N+Strep+Stop: 61.5 ng/μL Miraculin C+Strep+Stop: 78.5 ng/μL Brazzein N+Strep+Stop: 83.4 ng/μL Brazzein C+Strep+Stop: 99.4 ng/μL |