Protocol for the Taq Control PCR Reaction
Amplification of a 500 bp Lambda Fragment from the control mix. All kit components should be mixed and spun down prior to pipetting.
- Prepare the following 50 µl reaction in a 0.5 ml PCR tube on ice:
38.8 µl nuclease-free water
5 µl 10X Standard Taq Reaction Buffer
1 µl 10 mM dNTP Solution Mix
5 µl Control Template/Primer Mix
0.2 µl Taq DNA Polymerase*
* Due to the difficulties in pipetting small volumes of enzyme, Taq DNA Polymerase can be diluted in Diluent F (NEB #B8006S) or 1X reaction buffer. For example, 1 µl of Taq DNA Polymerase is mixed with 4 µl of diluent and 1 µl of that mixture is added to the reaction. Enzyme diluted in Diluent F can be stored at -20°C for future use.
- Gently mix the reaction and spin down in microcentrifuge.
If the thermocycler does not have a heated cover, add one drop of mineral oil to the reaction tube to prevent evaporation.
- Cycling conditions for Control PCR: