Keep everything cold all the time for better results
Protocol
Fill out the PCR excel spreadsheet and printout the sheet.
Sign-up for a PCR machine and make sure the correct PCR program exists on the machine.
Thaw DNA samples and controls and place on ice
Gather PCR cocktail ingredients (except for the taq) and PCR primers and put on ice. Make sure you have all the ingredients you need including sufficient amounts of primers.
Label PCR tubes or plates and place on ice in a 96 tube rack.
Aliquot DNA samples and controls (usually 1 or 2 µl containing between 100 to 500 ng DNA) into PCR tubes or plates. Touch pipette tip to bottom of tube when aliquoting, so DNA is at the bottom of the tube.
Mix the PCR cocktail using the amounts listed on the PCR excel spreadsheet in a 1.5 ml microcentrifuge tube. Always add the Taq polymerase last. Mix the complete cocktail by pipetting up and down. The Taq polymerase is normally in a glycerol solution, so it needs to be mixed well.
Aliquot the PCR cocktail into the PCR tubes or plates. If pipette tip touches a tube, throw it away and use a clean one.
Put caps on the PCR tubes or plates and seal completely.
Spin PCR tubes/plates.
Place PCR tubes/plates in PCR machine and start the PCR program.
Store PCR tubes/plates at 4ºC until they are analyzed on an agarose gel.