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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.