User:Meng Xiao He/Notebook/fall08/2008/11/19

Main project page Previous entry      Next entry

One primer Phusion PCR of suicide vectors extension time: 4:30 annealing temp: 52C primer: 1ul (in 25ul of rx) of M13F primer from TOPO kit cycles: 50 DNA (old minipreps of cbb, cco, Duo): 2ul each for control: TOPO A Confirmation of pure cultures Phusion PCR new TOPO cco, pER-cbb, pER-cco, pER-Duo#5, 7 colonies of TOPO C plate Rx: 5ul mastermix, 4.3ul water, 0.5ul primer pair, touch of DNA/cells Conditions: 2min initial denaturation at 98, 50s extension, annealing at 57C, 30 cycles Loss of plasmid Streaked out cultures of WM3064 on LB-DAP-agar, inoculate fresh LB-DAP liquid culture