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Avoid PCR Contamination

A How To Guide on How to Avoid PCR contamination.

PCR is a molecular biology technique which allows the production of more than 10 million copies of a target DNA sequence from only a few molecules of DNA. The sensitivity of PCR means that the sample used for PCR should not be contaminated with any other DNAs that may reside in the laboratory environment.

  • DNA Sample Preparation such as Genomic DNA preparation or Mini-Prep should be done in an area far or at least separate from the area where PCR reaction mixes will be prepared.
  • Laminar flow cabinet can be used and equipped with a UV lamp to prevent bacterial growth. This is the ultimate PCR reaction mixture work area.
  • Fresh Gloves should be worn at all times when PCR is performed.
  • Pipette tips with aerosol filters allow the prevention of microdroplets being injected into the PCR mixture, and thus prevent contamination of PCR reaction mixtures.
  • Negative Controls should be performed, in which the reaction mixture does not have the DNA template. If bands are seen after PCR, they are either contaminants or primer dimers.

Can't Find the Solution to your PCR Problem? Make sure you visit the PCR Forum for PCR troubleshooting and solutions to common PCR problems. You can also post questions to other researchers by simply registering first and then clicking on Post New Thread button in the forum, or simply clicking the question mark below!

PCR Forum Topics

loading dye Has anybody ever come across a loading dye that ran at a different molecular size on an agarose gel than expected? Why is this happening?
Cresol Red/Sucrose Hi. Does anyone have experience using cresol red/sucrose as loading dye added to the reaction BEFORE PCR? Does cresol red/sucrose have to have a...
Amplification won't work! I'm trying to amplify a ~200bp segment from a ~4kb plasmid. A postdoc in my lab previously amplified the segment and gave me a stock of gel-extracted...
Any ideas why my PCR amplification won't work? I'm trying to amplify a ~200bp segment from a ~4kb plasmid. A postdoc in my lab previously amplified the segment and gave me a stock of gel-extracted...
Heated lid on RFLP: is it ok? I'm doing three different RFLPs with three different restriction enzymes using a single amplicon, each RFLP in different tubes of course as each...
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