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

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

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