12 Things to rememeber while Setting up a PCR Reaction

This is a guest post by one of the blog readers.

• Consumables for setting up a PCR reaction such as PCR Tubes, Pipettes, PCR Racks should be cleaned with Hypo and then expose to UV for atleast 20 minutes.

• Before Setting up a PCR reaction Wash Hands,Wear Gloves.

• Always use Aerosol barrier tips (ART - Aerosol Resistant pipette Tips) for setting up PCR reaction, changing tips for each step.

• Use PCR grade water for diluting the stocks and for setting up a PCR Reaction.
  

• Use separate rooms for performing Mastermix preparation, Sample addition & Post PCR.

• Always use fresh aliquot of Reagents.

• Reagents for PCR should be thawed completely,vortex it and centrifuge before use.
  

• Never enter Post PCR room before setting up your Reaction.

• Accurate Pipette handling is must - Use calibrated pipettes.

• Start the PCR Reaction immediately after setting up the reaction this will help in avoiding Primer dimer formation.
  

• Positive and Negative controls should be kept in each PCR reaction without that PCR reaction is not considered as valid.

• Always perform PCR in Duplicates or in Triplicates.

• Make Sure you have cross checked the calculations before proceeding with the reaction.

Contamination in PCR lab is a Disaster....!!! Handle PCR Products (amplicon) Carefully.