Why is it necessary to purify PCR products before sequencing_FAQs_Information Center

Primers or RNA-contaminated samples will lead to misinterpretation of sequencing data. Thus, the purity of PCR products is the most critical factor for the success of sequencing because the contaminants with nucleases, salts, nucleotides, and ethanol decreases read length and signal intensity and even results in unreadable sequence data.

The goal of purifying products before sequencing is to remove the excess primers and preserve the ratio of the dNTP to ddNTP, which is necessary for efficient sequencing reactions. Classical methods used to clean up PCR products before sequencing include gel electrophoresis, ethanol precipitation, and column chromatography. But these methods can be not applicable when processing multiple samples. If more than one PCR product is present, column purification, ethanol precipitation, or enzymatic purification will not isolate the desired product. So it’s suggested to use gel purification to isolate the desired product or reoptimize the PCR to obtain a single product.

Leading Biology with an effective platform of PCR product preparation prior to sequencing is simple to use and ensure the result accuracy, which is a good solution compared to classical methods of PCR product clean-up prior to sequencing.

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Why is it necessary to purify PCR products before sequencing

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