User:Lu Wang/Notebook/Team Allergy/2010/06/30

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Procedures

Yesterday, we plated the allergen panel. If we have transformants for hpRNA today, then we will culture colonies and miniprep them for their DNA. Arabadopsis gDNA also arrived, so we will attempt to PCR out introns PAL and PME. We will also redo ligations for amiRNA, including a negative control.

hpRNA

  1. Culture colonies
  2. miniprep
  3. PCR arabadopsis introns PAL and PME

amiRNA

  1. redo ligation and transformation

Results

Culture Colonies Only Bet S worked, but there are only two colonies, so they may be background. We cultured a colony for each tube at 10am.

Miniprep The concentrations of plasmids after the miniprep are 80ng/μL and 108ng/μL

PCR for PAL and PME


Redo ligations and transformation Plated Bet, LTP, GFP, LTP 0.5 and GFP 0.5, as well as a negative control (just V0120 w/o insert added) at noon. We let the cells sit at room temp instead of in the incubator because we won't be able to check them until tomorrow morning.

Ligation Reactions
Bet GFP LTP LTP(.5) GFP(.5) Negative Control
DNA Insert 1 1 1 1 1 0
T4 DNA ligase buffer (10x) 1 1 1 1 11
diH2O 6 6 6 6 6 7
T4 DNA ligase 1 1 1 1 1 1
DNA Backbone 1 1 1 1 11



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