3. How to ensure laboratory detection quality for PCR experiments?
2026-03-24 11:17
1) A PCR laboratory should have sufficient and reasonable space, good lighting and air conditioning equipment. Although the working environment is not a direct factor of detection quality, a spacious and comfortable laboratory environment is certainly conducive to quality assurance;
2) Manage instruments and equipment reasonably and effectively with regular maintenance and calibration to keep them in good condition (e.g., calibrate pipettes regularly for sufficient accuracy and precision; maintain the optical system of real-time fluorescent PCR instrument and temperature difference between reaction wells within allowable ranges; also include the management of balances, centrifuges, refrigerators, etc.);
3) Select ideal reagents by controlling internal factors (sample processing methods, raw materials for nucleic acid amplification, methodological design, etc.) and external factors (kit transportation and storage conditions);
4) Establish standardized operating procedures: A complete PCR process includes sample collection, transportation, storage, numbering, reagent preparation, nucleic acid extraction, amplification, product detection, result analysis and reporting. Formulate scientific laboratory-specific SOP documents and implement them strictly;
5) Identify major pollution sources and take prevention measures: Main PCR pollution sources include a large number of target microorganisms in samples, cloned plasmids, specific microorganisms in the laboratory and residual contamination from previous amplification products (the main causes of false positives). Strictly partition the laboratory, follow the workflow, clean workbenches with chemical agents, and eliminate amplification product contamination by UV irradiation and UNG method;
6) Conduct internal and external quality control: Internal quality control ensures the consistency of in-lab testing results; external quality control provides data for retrospective comparison of laboratory testing results with objective inter-laboratory standards.
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