PCR Amplification with ProtoScript® M-MuLV Taq RT-PCR Kit


We recommend 2–5 μl of the diluted cDNA product per 50 μl PCR reaction.


  1. Mix the following in a PCR tube on ice:
    Taq 2X Master Mix 25 μl (mix well by inverting before use)
    Forward Primer (10 μM) 1 μl (final concentration 200 nm)
    Reverse Primer (10 μM) 1 μl (final concentration 200 nm)
    Diluted cDNA 2–5 μl
    H2O variable
    Total Volume 50 μl
  2. Mix gently. Overlay with mineral oil if the thermal cycler lacks a heated lid.
  3. The following PCR cycling conditions are recommended for 0.2 ml thin-wall PCR tubes on Bio-Rad iCycler or similar thermocyclers.
    25–35 Cycles 94°C 30 seconds
    45–68°C 10-30 seconds
    68°C 1 minute per kb
    Final Extension 68°C 5-10 minutes
  4. Analyze 5 μl of the PCR product by agarose gel electrophoresis.