Best way to avoid PCR contamination
Separate Pre-PCR (clean area) and post PCR (dirty area)
PCR contamination can be a major problem in many laboratories. Here are some strategies to avoid PCR contamination:
1. Use separate workstations for pre- and post-PCR steps: Be sure to divide your work area and perform pre-PCR activities, such as DNA extraction and PCR setup, in separate rooms or areas to those where post-PCR processes such as PCR product analysis are being carried out.
2. Use dedicated equipment: Use new and sterile pipettes, filter tips, and microcentrifuge tubes for each PCR reaction. Have dedicated equipment such as thermal cyclers, electrophoresis chambers, and gel documentation systems for PCR and post-PCR processes.
3. Practice good laboratory hygiene: Use sterile gloves and lab coats. Clean all surfaces, equipment, and materials that come into contact with samples with DNA decontaminants, such as bleach.
4. Use PCR inhibitors: Use reagents that contain PCR inhibitors, such as BSA, when appropriate for your samples.
5. Limit amplification cycles: Use as few PCR amplification cycles as possible to minimize the chances of contamination building up in subsequent cycles.
6. Avoid aerosol contamination: Use proper containment when handling sample materials or working with amplified DNA. Avoid aerosol generation during pipetting and be sure to keep your tubes and plates covered during amplification.
Overall, the best way to avoid PCR contamination is to be vigilant in all aspects of laboratory practice. Being proactive in creating separate workstations, using dedicated equipment, employing proper hygiene, using PCR inhibitors, and avoiding aerosol contamination can help prevent contamination build-up in both pre- and post-PCR steps.
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