Boosting PCR accuracy – tips for maximizing amplification fidelity

S3E12 - Boosting PCR accuracy – tips for maximizing amplification fidelity

Published: June 25, 2025

Duration: 18:41

In this Mol Bio Minutes episode, Laurynas Alijošius shares a personal story that every molecular biologist can relate to—running PCR, cloning, and sequencing, only to discover frustrating errors in the DNA. 

This episode dives deep into PCR accuracy and why it matters for everything from sequencing to cloning and long-read library prep. Laurynas breaks down the major contributors to PCR error, including the fidelity of DNA polymerase, primer design flaws, template impurities, and suboptimal cycling conditions. He then offers a range of solutions—like switching to high-fidelity enzymes, using ready-to-go master mixes, and optimizing magnesium ion co...