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Old 06-18-2015, 09:57 PM   #5
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,232
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That is less likely. For end repair (filling in 5 overhang and removing 3 overhang) two enzymatic activities are required so it is less likely to add 3 A while another enzyme is removing 3 overhangs. This can be tested by omitting incubation at 65C and immediately cleaning up end repair reaction followed by adapter ligation. Lack of 3 A should significantly reduce ligation efficiency.
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