An Efficient Polymerase Chain Reaction (PCR) Enhancer for Highly Guanine-Cytosine (GC)-Rich DNA Sequences

Abstract

Background: Polymerase chain reaction (PCR) has become a fundamental technique in molecular biology. Nonetheless, PCR amplifications are frequently impaired by high GC content of the target sequence, leading to low yield and specificity of products, with no product at all in the worst cases. Locally high-temperature melting regions within the template can act as permanent termination sites.Method: Here we designed and tested an effective and low-cost PCR enhancer, a combination of dimethyl sulfoxide (DMSO) 10% (v/v) and magnesium chloride (MgCl2) 1,5 mM that broadly enhanced the qualitative output of PCRs. We used Mycobacterium tuberculosis strain H37vR as a PCR template.Result: It was found that PCR enhancer containing 10% (v/v) of DMSO and 1,5 mM of MgCl2 improved the amplification of GC-rich template of M. tuberculosis gene other than without the PCR enhancer.Conclusion: Therefore, this PCR enhancer could be widely useful to improve the amplification of GC rich construct from another genome.