User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/07

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09/07/14

  • Minipreps & digests

Minipreps

  • Used Sigma kit (ran out of lysis buffer)
    • Resuspended 4 mL cell culture (pelleted) in 400 μL resuspension buffer
    • Lysed with 60 μL Zymo 7x blue buffer
    • Followed rest of Sigma protocol


Digests
> Check with KpnI digests (same as 9/14/10, Harvard)

Reagent Volume Expected:
Backbone = 7489
Empty vector frag. = 913
1. KAH160/KAH015-1,2,3 = 2060
2. KAH165/KAH015 = ---
3. KAH170/KAH015-1,2,3 = 1868
File:KAH 091410 gel1.tif
15 μL/lane; 1% agarose
DNA(plasmid) 1.0 μL
10X buffer 1.5
KpnI 1.0
dH2O 11.5
  15 μL --> 37°C/ ~15 min.
 
 
 
 

--> Reaction conditions