IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/08/03

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Team Allergy
ladder,
 * Grew up cultures of completed ihpRNA in pORE expression vector (Bet2, LTP, Ger)
 * amiRNA PCR
 * PCR for each of the seperate parts seemed to have work

Team Fence
VP16 PCR from last night

Yesterday's transformations




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 DH2O

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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