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

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Contents

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 polymerase2x5
MilliQ3.5
total10
98C30 sec
98C10 sec
65C30 sec
72C30 sec
30 cycle
72C5 min

Load 2.5ul sample

image:0062.jpg

Fragments extraction

E-Gel 2% 15 min

Image:0063.jpg

18 min

Image:0064.jpg

Bisulfite conversion

Reaction mixture 1x
Sample20
Bisulfite Mix85
DNA Protect Buffer35
total140
95C5 min
60C25 min
95C5 min
60C85 min
95C5 min
60C175 min
20Co/n


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