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. Author manuscript; available in PMC: 2012 Oct 23.
Published in final edited form as: Methods Mol Biol. 2012;882:431–468. doi: 10.1007/978-1-61779-842-9_25

Table 9.

Composition of reaction master mix for Expand Long Template PCR Reaction

Components Volume in Each Reactiona
10X Expand Long Template Buffer 3 5 µl
dNTP (10mM) 2.5 µl
Forward primer (10 µM) (Table 2) 1.5 µl
Reverse primer (10 µM) (Table 2) 1.5 µl
Expand Long Template Enzyme mix 0.75 µl
Template DNA Added at later step
Water Bring final volume including DNA to 50 µl
a

The volume for a single reaction is 50 µl so multiple the number of amplification reactions desired by 50 to determine how much reaction master mix to make. Always make more than you need to account for losses during pipetting.