Haynes Lab:Notebook/Jan/2015/03/27

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03/27/15

  • Cultures - BD006/MV2 colonies (from 3/26/15 ligations), 5 mL each (4 total)
  • Plasmids - DPRE minipreps (5 mL cultures)



Plasmids - DPRE minipreps

  • ---Karmella 15:12, 27 March 2015 (EDT):
  • Sigma GenElute Miniprep kit.
  • Elute with 70μL elution buffer
  1. DPRE a
  2. DPRE b
  • Digests (Fermentas FD)
    • E/S to prep for insertion upstream of BD004
    • Also digest BD004 w/ E/X
Reagent Volume Expected:
1,2. DPRE (E/S) = 1740*, 3200
3. BD004 (E/X) = 7316*
*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
    • All look good


  • Miniprep conc.'s
Sample OD260 260/280 ng/μL
1. DPRE a mp 0.411 1.89 411.47
2. DPRE b mp 0.411 1.89 410.51


  • Gel purification conc.'s
    • Sigma gel extraction kit, eluted & back-eluted w/ 30 μL elution sln
Sample OD260 260/280 ng/μL
1. DPRE a (E/S) 0.017 1.88 16.81
2. DPRE b (E/S) 0.013 1.54 13.41
2. BD004 (E/X) 0.006 1.85 6.46