Lyophilized Ready to Use & Load PCR Master Mix
- Add 450 uL of PCR quality water to the tube containing dried mix for 25 reactions of 20 uL each.
- Completely dissolve the powder by gently and briefly vortex, touch spin the tube before open the tube.
- Aliquot the needed volume of Ready to Use & Load PCR Mix into a 0.5 mL tube, 18.5 uL per PCR reaction (e.g., 5 reactions = 92.5 uL).
- Add 5 pmol of each PCR primer (e.g., 0.5 uL of a 10 pmol/uL stock) (e.g., 5 reactions = 2.5 uL each primer).
- Briefly vortex to mix primers and the Ready to Use & Load PCR Mix, touch spin the tube before open the tube.
- Aliquot the solution into PCR tubes (19 uL each).
- Add 1 uL of the DNA sample.
- Mix the reaction by briefly vortex the PCR tubes in the rack.
- Touch spin the PCR tubes, run cycling program in a thermocycler (the reaction volume is 20 uL).
- Load 3-10 uL of each reaction directly to an agarose gel containing DNA staining dye (eg. casted in TBE buffer containing 0.5 ug/mL (gel) ethidium bromide or other DNA staining dye).
- Stop the gel run when the red tracking dye has reached an appropriate location on the gel for your sample (see photo below).
- The gel can be directly viewed and photographed on a UV table or other light source or a fluorescence scanner according to the DNA dye used.
- Any remaining Ready to Use & Load PCR Mix can be kept on ice for use the same day or freeze at -20 °C for a later use. Repeated freeze-thaw may decrease activity.