Table 9.
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 |
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.