Template:SBB-Protocols EcoBamTransfer

EcoRI/BamHI Transfer
1. Set up digest: For BBb transfers: 5 uL ddH2O 1 uL NEB2+ATP Buffer 0.5uL EcoRI 0.5uL BamHI 3 uL miniprepped plasmid
 * Set up the digest (you can do mastermixes of the water/buffer/enzymes):
 * run temperature program on thermocycler of 1hr at 37 and then 20 min at 80

2. Add 0.5uL of predigested vector and 0.5uL of T4 DNA ligase 3. Transform into correct cell line 4. Rescue for 45min to 1hr in 37C shaker 5. Plate on correct antibiotics
 * Incubate 30 min. at room temp