Home Protocols LongAmp™ Taq 2X Master Mix Protocol (M0287)

LongAmp™ Taq 2X Master Mix Protocol (M0287)

Protocol

  1. Thaw on ice and mix well by inverting several times before use.
  2. 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.
  3. Transfer PCR tubes to a PCR machine with the block preheated to 94-95°C and begin the programmed cycling:

    STEP
    		TEMP
    		TIME
    Initial Denaturation
    		95°C
    		30 seconds
    25-45 Cycles 95°C
    50-65°C
    65°C
    		10 seconds
    10-60 seconds
    50 seconds per kb
    Final Extension 65°C
    		10 minutes
    Hold 4-10°C