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. Author manuscript; available in PMC: 2010 Apr 22.
Published in final edited form as: Nat Protoc. 2009 Sep 17;4(10):1471–1501. doi: 10.1038/nprot.2009.98
Component Amount Final Concentration
Purified PCR product of step 49 24 μl unknown
Forward primer OK1430 (10 μM) 3 μl 0.6 μM
Reverse primer OK 1432 (10 μM) 3 μl 0.6 μM
10× Expand Buffer with MgCl2 5 μl 1×
10mM dNTPs 4 μl 0.8 mM
Nuclease-free water 10.625 μl
Expand Enzyme (3.5 U μl-1) 0.375 μl 0.02625 U μl-1
Total 50 μl