Haynes Lab:Notebook/Jan/2015/03/28

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03/28/15

  • Minipreps - BD006/MV2 colonies (from 3/26/15 ligations), 5 mL each (4 total)



Minipreps - BD006/MV2 colonies

  • Sigma GenElute kit.
    • Elute & back-elute w/ 70 μL elution sln.
  1. BD006/MV2 plate 1, a
  2. BD006/MV2 plate 1, b
  3. BD006/MV2 plate 2, a
  4. BD006/MV2 plate 2, b


  • Restriction Digest check (Fermentas FD)
    • Cut with ApaLI (E/S insertion got rid of BB RFC23 cut sites)


Reagent Volume Expected:
* BD006/MV2 = 7207, 1243, 288
* MV2 only = 2998, 1243, 288
* BD006/V0120 = 7036, 497
Hover name
30 μL/lane, 1% agarose; Ladder
DNA (plasmid) 3.0 μL
10x buffer 1.5
ApaLI 1.0
dH2O 9.5
  15 μL --> 37°C/ ~10 min.


  • Conclusions
    • Success! All four look like BD006/MV2
    • Submit for sequencing next week


  • Miniprep conc.'s
Sample OD260 260/280 ng/μL
1. BD006/MV2 1-a 0.274 1.44 274.10
2. BD006/MV2 1-b 0.299 1.93 298.68
3. BD006/MV2 2-a 0.331 1.93 330.69
4. BD006/MV2 2-b 0.306 1.90 305.94