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Old 05-27-2015, 10:52 AM   #4
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Location: Bay Area

Join Date: Jun 2012
Posts: 120

I believe the main reason you typically see cleanup between 2nd strand and end repair is that the 2nd strand buffer for most systems has ammonium sulphate in it, and the T4 PNK used in end repair is potently inhibited by ammonium. It might work out if the reaction is diluted enough that the ammonium is a nonfactor.
Ex, NEBNext
20 mM Tris-HCl
12 mM (NH4)2SO4
5 mM MgCl2
0.16 mM β-NAD
0.19 mM dNTPs each
pH 7.4 @ 25C
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