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PCR Contamination

PCR Contamination - PCR - Polymerase Chain Reaction Forum

PCR Contamination - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 11-04-2007, 01:46 PM
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Question PCR Contamination



How can one minimize PCR contamination?

I have tried many times but I keep getting extra bands.

Where do you start? primers? water? buffers?

thanks
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Old 11-04-2007, 06:04 PM
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Default Re: PCR Contamination

So many tips for minimizing PCR contamination:

Lab setup:
Have separate rooms for sample storage, extraction, PCR reaction preparation, and electrophoresis.
Keep all reagents in separate freezer from samples.
Always wipe down all working surfaces.
Avoid aerosolizing any DNA/RNA.

PCR:
Use different sets of pipettors for extraction and PCR reaction setup.
Work under a PCR hood that is decontaminated by UV on daily basis.
Use aerosol-barrier tips.
Keep primers and enzymes in small aliquots so you replace them often.

If you have contamination:
Dump as many of your cheap reagents as possible.
Get acquainted with the PCR troubleshooting flow chart ---> [Only registered users see links. ]
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