Buckner Lab:Notebook/Summer 2010/2010/05/18

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Plasmid Isolation and Liquid Media

Objectives:

  • Analyze bacterial cultures inoculated 5/17
  • Prepare LB Broth medium for the Synthetic Biology Lab
  • Make a ampicillin stock for the SynBio Lab.
  • Make overnight cultures

Bacteria Results

fas1b 2, KER 1D, Kn1 Recombined Plasmids in E.coli JM109
Bacterial Strain Growth (+/-) Description
JM 109 25μL + Restreak (MC)
JM 109 100μL + Good Growth (MC)
Kn1 25μL + Sm iso colonies
Kn1 100μL + MC to isolate
KER 1D 25μL + Sm iso colonies
KER 1D 100μL + MC to isolate
fas1b 2 25μL + iso colonies
fas1b 2 100μL + MC to isolate

MC= "multiple colonies present on agar"

LB Broth and Ampicillin Stock

-Made 300mL of LB broth media. Autoclaved at 9:47am
-Poured broth into 6, 50mL portions. Stored in SynBio Lab at room temp
-Started to make 10mL Amp (50mg/mL concentration) but spilled some during sterilization. Ended making 6mL of Amp stock and separating into 7 x's 700μL portions. Placed sterilized Amp in bullet box in Buckner freezer.


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