Haynes Lab:Notebook/Jan/2015/04/07

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04/07/15

  • Minipreps - BD004 assembly



Minipreps - BD004 assembly

  • ---Karmella 17:57, 7 April 2015 (EDT):
  • Sigma GenElute, minipreps from 2x 5 mL cultures
  • Elute & back-elute w/ 70 μL elution sln.
  1. BD004 a
  2. BD004 b


  • Digests (Fermentas FD)
    • E/S to prep for insertion into MV2 (E/X)


Reagent Volume Expected:
1,2. BD004 (E/S) = 4096*, 3200
*Bands cut & purified
Hover name
30 μL/lane, 1% agarose; Ladder
DNA (plasmid) 20.0 μL
10x buffer 3.0
enzyme 1 1.0
enzyme 2 1.0
dH2O 5.0
  30 μL --> 37°C/ ~30 min.


  • Conclusions
    • tba


  • Miniprep conc.'s
Sample OD260 260/280 ng/μL
1. BD004 a mp 0.357 1.92 356.67
2. BD004 b mp 0.262 1.89 262.29


  • Gel purification conc.'s
    • Sigma gel extraction kit, eluted & back-eluted w/ 30 μL elution sln
Sample OD260 260/280 ng/μL
1. BD004 a (E/S) 0.032 1.73 32.13
2. BD004 b (E/S) 0.02 1.62 19.79




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