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.
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