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BioTechniques

Improving sequencing quality from PCR products containing long mononucleotide repeats.


PMID 20569204

Abstract

Stutter products are a common artifact in the PCR amplification of frequently used genetic markers that contain mononucleotide simple sequence repeats. Despite the importance of accurate determination of nucleotide sequence and allele size, there has been little progress toward decreasing the formation of stutter products during PCR. In this study, we tested the effects of lowered extension temperatures, inclusion of co-solutes in PCR, PCR cycle number, and the use of different polymerases on sequence quality for a set of sequences containing mononucleotide A/T repeats of 10-17 bp. Our analyses showed that sequence quality of mononucleotide repeats

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