User:Karmella Haynes/Notebook/BioBrick cloning/2011/01/13

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01/13/11

  • ✓ Minipreps: KAH171(2), KAH172(2), KAH173(6), MI127(2)
  • ✓ Sequencing order: KAH171, 172


Minipreps
> Check KAH173 mp's with E/P digests

Reagent Volume Expected:
1-6. KAH173 = 2960
V0120 = ~3200
KAH73 = 2438
HPK = 516 		1/13/11 E/P digest
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.

--> Conclusion: Not confident about these, especially since the ratio was ~1:2. Re-do cloning. Dephos. vector and use more insert.

Sequencing (Genewiz, standard)

  1. KAH171-1, P0001: good (keep)
  2. KAH171-2, P0001: good
  3. KAH172-1, P0001: good (keep)
  4. KAH172-2, P0001: good



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