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

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Team Allergy

  • 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

[[Image: |240px|PCR gel]] ladder,

Team Fence

VP16 PCR from last night File:83gelpcrvp16.jpg

Yesterday's transformations

Gal4-EcR lig 8-3.jpg EcR neg control 8-3.jpg LTP lig-1 8-3.jpg LTP lig-2 8-3.jpg B11 neg cont lig 8-3.jpg

Barn-strep lig 8-3.jpg B15 cont lig 8-3.jpg

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
    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