User:Karmella Haynes/Notebook/BioBrick cloning/2009/12/04

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12/04/09

  • ✓ Minipreps: KAH93(2), 94(2), 95/MV2(2)
  • ✓ Cultures: KAH99(2), 100(2), 101(2), 105(2), 106(2), 107(2), 108/MV2(2)
  • ✓ Check sequencing results for KAH011 (NLS-myc-stop)

Minipreps
> Check with E/P/ScaI digests

Reagent Volume Expected:
1,2. KAH93/MV2 = 4839
3,4. KAH94/MV2 = 5011
5,6. KAH95/MV2 = 5083
MV2 fragments = 3811, 709
File:KAH 120409 gel1.tif
15 μL/lane; 1% agarose
Confirm by sequencing. Pick 2 more clones for KAH95/MV2.
DNA(plasmid) 2.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
ScaI 1.0
dH2O 8.5
  15 μL --> 37°C/ ~15 min.

Sequencing results

  1. KAH011-1: bad clone
  2. KAH011-2: good ✓ (Keep)
  3. KAH011-3: good ✓
  4. KAH011-4: good ✓
  5. KAH011-5: good ✓
  6. KAH011-6: good ✓
  7. KAH011-7: good ✓
  8. KAH011-8: good ✓