User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/04/06

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Apis mellifera methylome

Confirmation of the adapter ligation

Reaction mixture conc. 1x
Callow (after adapter ligation, before pulification) ?? 0.5
PCR primer PE 1.0 ?? 0.5
PCR primer PE 2.0 ?? 0.5
Phusion DNA polymerase 2x 5
MilliQ 3.5
total 10
98C 30 sec
98C 10 sec
65C 30 sec
72C 30 sec
30 cycle
72C 5 min

Load 2.5ul sample

0062.jpg

Fragments extraction

E-Gel 2% 15 min

0063.jpg

18 min

0064.jpg

Bisulfite conversion

Reaction mixture 1x
Sample 20
Bisulfite Mix 85
DNA Protect Buffer 35
total 140
95C 5 min
60C 25 min
95C 5 min
60C 85 min
95C 5 min
60C 175 min
20C o/n