A | B | C | |
Reagent | Final concentration | Volume per reaction | |
Nuclease-free water | - | 6.32µl | |
Platinum II Taq | 0.04U/µl | 0.08µl | |
5x Platinum II Buffer | 1x | 2µl | |
10mM dNTP | 0.2mM each | 0.2µl | |
10uM F | 0.2uM | 0.2µl | |
10uM R | 0.2uM | 0.2µl | |
DNA* | - | 1µl | |
10µl reaction |
A | B | C | D | |
Initial denaturation | 94°C | 2 min | ||
Denature | 98°C | 5 sec | ||
Anneal and extend | 60°C | 15 sec | To step 2 x25 | |
Hold | 4°C | ~ |