User:Alexander S Mikheyev/Notebook/bee methylome/2010/04/14

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Library preparation Library amplification Reaction mixture conc. Callow Mature template (after bisulfite conversion) ?? 2.5 5 PCR primer PE 1.0 ?? 0.5 0.5 PCR primer PE 2.0 ?? 0.5 0.5 Phusion DNA polymerase 2x 12.5 12.5 MilliQ 9 6.5 total 25 25 98C 30 sec 98C 10 sec 65C 30 sec 72C 30 sec 25 cycle 72C 5 min load 1.0ul of reaction Gel extraction lane ' 1 Callow 2 empty 3 empty 4 2log ladder (NEB) 3.0 ug 5 100 ladder (TaKaRa) 1.0 ug 6 empty 7 empty 8 Mature M 1kb plus DNA ladder (Invitrogen) 2.0 ug load all (E-Gel 2.0%) Followed by purification using QIAquick PCR purification kit