Author: Warnecke Peter M. Stirzaker Clare Melki John R. Millar Douglas S. Paul Cheryl L. Clark Susan J.
Publisher: Oxford University Press
ISSN: 1362-4962
Source: Nucleic Acids Research, Vol.25, Iss.21, 1997-11, pp. : 4422-4426
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Abstract
Methylation analysis of individual cytosines in genomic DNA can be determined quantitatively by bisulphite treatment and PCR amplification of the target DNA sequence, followed by restriction enzyme digestion or sequencing. Methylated and unmethylated molecules, however, have different sequences after bisulphite conversion. For some sequences this can result in bias during the PCR amplification leading to an inaccurate estimate of methylation. PCR bias is sequence dependent and often strand-specific. This study presents a simple method for detection and measurement of PCR bias for any set of primers, and investigates parameters for overcoming PCR bias.
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