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Effect of single nucleotide mismatch at 3′-end of the primers in selected candidate micro RNA breast cancer genes

By: Aftab Ali Shah, Mushtaq Ahmed, Farman Ullah

Key Words: Breast cancer, Micro RNA, PCR, Hsa-mir-146a, Hsa-mir-196a-2.

Int. J. Biosci. 9(1), 430-434, July 2016.

DOI: http://dx.doi.org/10.12692/ijb/9.1.430-434

Abstract

Polymerase chain reaction (PCR) is an important biological tool for qualitative and quantitative detection and amplification of desired sequence of DNA (deoxyribonucleic acid) in living organisms. The success of PCR reaction is based on the specificity of the primers used. The potential of these primers can be influenced by primer-template DNA, primer-primer complementarily, annealing, extension temperatures and nature of the mismatch. But, often the powers of thermal cycler are restricted due to primer–template mismatches which can lead to inaccuracies/false results. In the present study, the effect of single nucleotide mismatches (No change, T/A and C/G) at the 3′-end of primers on PCR reaction of breast cancer candidate microRNA genes i.e; hsa-mir-196a-2 (1), hsa-mir-196a-2 (2) and hsa-mir-146a respectively was evaluated in the human genome using conventional PCR. Here we conclude that no effect was found on the PCR product of these studied genes with introduced single mismatches at the 3′-end of primers under standard PCR conditions.

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