Haynes Lab:Notebook/Synthetic Chromatin for Cancer Research/2015/06/17

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06/17/15

Minpreps & Digests

  • Minipreps
    • Sigma GenElute kit, elute w/ 50 μL sterilized water.
  • Digests
    • Check with E/S digests; cannot use PstI because PLrigid & PLrigid4 contain PstI sites
Reagent Volume Expected:
1. DT009 = 2668, 252
Empty PLflex = 2668, 95
2. DT010 = 2656, 432
Empty PLflex4 = 2656, 274
3. DT011 = 2668, 252
Empty PLrigid = 2668, 95
4. DT012 = 2656, 432
Empty PLrigid4 = 2656, 275
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 2.0 μL
10X buffer 1.5
EcoRI 1.0
SpeI 1.0
dH2O 9.5
  15 μL --> 37°C/ ~15 min.

CONCLUSIONS:

  1. DT009 - success.
  2. DT010 - success.
  3. DT011 - success.
  4. DT012 - success.


  • Next step:
    • Sequence verify all clones (extra precaution; linker part is small)



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