Haynes Lab:Notebook/Jan/2015/04/10

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04/10/15

  • Minipreps - BD004/MV2 (6x 5mL cultures)



Minipreps - BD004/MV2

  • ---Karmella 20:52, 10 April 2015 (EDT):
  • Sigma GenElute kit.
    • Elute & back-elute w/ 70 μL elution sln.
  1. BD004/MV2 a
  2. BD004/MV2 b
  3. BD004/MV2 c
  4. BD004/MV2 d
  5. BD004/MV2 e
  6. BD004/MV2 f


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


Reagent Volume Expected:
* BD004/MV2 = 7108, 1243, 288
* MV2 only = 2998, 1243, 288
* BD004/V0120 = 5659, 1246, 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, and very lucky! Clone "a" is the desired plasmid.
    • Keep clone a and discard other minipreps.
    • Mark the streak library accordingly.


  • Miniprep conc.'s
Sample OD260 260/280 ng/μL
1. BD004/MV2 a 0.400 1.90 399.66
2. BD004/MV2 b 0.344 1.90 344.33
3. BD004/MV2 c 0.271 1.88 270.79
4. BD004/MV2 d 0.292 1.89 292.47
5. BD004/MV2 e 0.279 1.90 279.28
6. BD004/MV2 f 0.221 1.88 220.53