Sources of PCR inhibitors
Mechanism of action of PCR inhibitors
Removal of PCR inhibitors1. The most fundamental way to overcome PCR inhibition is to remove the inhibitor from the sampleA. Sample processing For different types of samples, appropriate pretreatment methods such as digestion, dissolution, washing, etc. are used to remove inhibitors. (1) Plant samples: Extraction methods such as CTAB and SDS were used to remove inhibitors such as polyphenols and polysaccharides. (2) Animal samples: Inhibitors such as proteins and lipids can be removed by methods such as proteinase K digestion and phenol-chloroform extraction. (3) Microbial samples: Reagents such as sodium chloride and SDS can be used, combined with centrifugation and washing steps, to remove inhibitors such as humic acid. B. Sample Purification Use purification methods such as filtration columns and magnetic beads to remove inhibitors. Second, the simplest way to overcome PCR inhibition is to dilute the sample, thereby reducing the concentration of PCR inhibitors. 3. The most worry-free method is to use a modified DNA polymerase or reaction system that is resistant to PCR inhibitors. For example, by using enzymes with strong tolerance to inhibitors or adding stabilizers such as bovine serum albumin (BSA) to the reaction system, the success of subsequent analysis can be guaranteed. However, the cost of improved enzymes is often high, and different improved enzymes and optimized reaction systems may be required for different PCR inhibitors. To ensure optimal experimental results, while removing PCR inhibitors, we should also avoid introducing new PCR inhibitors. For example, when selecting reagents, we should choose high-quality, inhibitor-free enzymes, dNTPs, and primers. During the experiment, we should strictly follow the experimental protocol to ensure that no inhibitors are introduced. We should also ensure that the experimental equipment is thoroughly washed to avoid cross-contamination. SummarizePCR inhibitors are the "invisible killers" that can hinder the success of PCR experiments. Understanding their sources, mechanisms of action, and removal methods is crucial for conducting PCR experiments. By implementing a series of measures, such as optimizing sample handling, selecting high-quality reagents, and standardizing experimental procedures, we can minimize the impact of inhibitors and ensure the smooth progress of experiments. Related products
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