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==Protocol== ===PCR amplification and preparation for sanger sequencing=== *Add 12.5ul of 100ul of F and R primer and add 475ul sterile h2o to make 2.5uM primer mix {| {{table}} | align="center" style="background:#f0f0f0;"|'''Content''' | align="center" style="background:#f0f0f0;"|'''Volume per reaction''' | align="center" style="background:#f0f0f0;"|'''MM''' |- | DNA||Pick one colony per well||Do not add to MM |- | H2O||25||205 |- | M13-21 F+R Primer (2.5uM each)||8||65.6 |- | 2X Taq MM||15||123 |- | Total||48||393.6 |} *Add 48ul MM to each colony 94C 10 min to break up the cells 94C 30 sec -> (94C for 30 sec, 60C for 30 sec, 72C for 1 min) x30 -> 72C 5 min ->15C forever ===PAGE=== [[File:ZhangLab_2 2011-08-19 16hr 58min.jpg|500px]]
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