Template:SBB-EcoBamXfers

From OpenWetWare

Revision as of 12:53, 2 February 2010 by JCAnderson (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search

Eco/Bam Transfer

Set up digest:

  • Set up the digest:
 For BBb transfers:
 7.5uL ddH2O
 1 uL NEB2+ATP Buffer
 0.5uL EcoRI
 0.5uL BamHI
 0.5uL miniprepped plasmid
  • run temperature program on thermocycler of 1hr at 37 and then 20 min at 80
  • Add 0.5uL of predigest vector and 0.5uL of T4 DNA ligase
  • Incubate 30 min. at room temp, transform, rescue, plate.
Personal tools