User:Alexander S Mikheyev/Notebook/bee methylome/2010/04/18

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Library preparation

' lane 1 2
Reaction mixture conc. Mature Mature (2nd PCR)
template (after bisulfite conversion) ?? 8.5 1
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 3 10.5
total 25 25
98C 30 sec
98C 10 sec
65C 30 sec
72C 30 sec
25 cycle
72C 5 min
Gel extraction by QIAGEN Gel extraction kit
followed by MinElute kit

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