User:Karmella Haynes/Notebook/BioBrick cloning/2010/03/23

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03/23/10

  • ✓ Minipreps: KAH142/V0201 (4)
  • ✓ Cultures: KAH143 (4)
  • ✓ Sequencing: KAH86, 96, 142/V0201



Minipreps
> Check with XbaI digest

Reagent Volume Expected:
Vector = 4516
1-4. KAH142/V0201 = 4571 (5212)
XbaI digest 3/23/10
15 μL/lane; 1% agarose
DNA(plasmid) 2.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 9.5
  15 μL --> 37°C/ ~15 min.

Sequencing (Genewiz, same day)

  1. KAH86 P0001: mistake, reverse primer (re-do)
  2. KAH96 P0001: mistake, reverse primer (re-do)
  3. KAH142/V0201 3T3: good, but did not read into promoter
  4. KAH142/V0201 BGHR: good

--> Re-submit KAH96 and 86 for sequencing (both directions, same day on Thurs.). If KAH96 is missing the promoter, need to re-build KAH142.


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