LongAmp™ Taq 2X Master Mix Protocol (M0287)
- Thaw on ice and mix well by inverting several times before use.
- Add the following components to a thin-walled PCR tube on ice:
Component 25 µl reaction 50 µl reaction Final Conc. LongAmp Taq 2X Master Mix 12.5 µl 25 µl 1X 10 µM Forward Primer 1 µl 2 µl 0.4 µM (0.05–1 µM) 10 µM Reverse Primer 1 µl 2 µl 0.4 µM (0.05–1 µM) Template DNA variable variable <1000 ng Nuclease-free water to 25 µl to 50 µl
Notes: Gently mix the reaction. Avoid pipetting samples containing target DNA when amplicons above 20 kb are desired. Collect all liquid to the bottom of the tube by a quick spin if necessary. Overlay the sample with mineral oil if using a PCR machine without heated lid.
- Transfer PCR tubes to a PCR machine with the block preheated to 94-95°C and begin the programmed cycling:
25-45 Cycles 95°C
50 seconds per kb
Final Extension 65°C