Bisulfite conversion-specific and methylation-specific PCR: a sensitive technique for accurate evaluation of CpG methylation.

Masahiro Sasaki; Jason Anast; William Bassett; Toshifumi Kawakami; Noriaki Sakuragi; Rajvir Dahiya

doi:10.1016/j.bbrc.2003.08.005

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Bisulfite conversion-specific and methylation-specific PCR: a sensitive technique for accurate evaluation of CpG methylation.

Masahiro Sasaki·Jason Anast·William Bassett·Toshifumi Kawakami·Noriaki Sakuragi·Rajvir Dahiya

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Abstract

Methylation-specific PCR (MSP) is frequently used to distinguish methylated alleles in the genome. Sequences that have been incompletely converted during bisulfite treatment are frequently co-amplified during MSP. For accurate MSP, it is important to detect methylated sequences in a background of unconverted DNA with a high level of sensitivity. We report here sensitive techniques, bisulfite conversion-specific MSP (BS-MSP) to accurately evaluate CpG methylation. BS-MSP provides accurate results across a wide spectrum of bisulfite conversion levels. BS-MSP is also confirmed to be a useful technique for the routine analysis of clinical tumor specimens that were paraffin-embedded and microdissected. BS-MSP thus provides the powerful features of ease of use and compatibility with paraffin sections. We recommend that methylation analysis should include a step to eliminate unconverted DNA to avoid overestimation of the DNA methylation level in the samples.

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Cadherins CpG Islands DNA Methylation DNA, Neoplasm Polymerase Chain Reaction Reference Values Reproducibility of Results Restriction Mapping Sensitivity and Specificity Sequence Analysis, DNA Sulfites Urinary Bladder Neoplasms Comparative Study Evaluation Study Journal Article Research Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.Validation Study

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Masahiro Sasaki, Jason Anast, William Bassett, Toshifumi Kawakami, Noriaki Sakuragi, Rajvir Dahiya. 2003-09-19. Bisulfite conversion-specific and methylation-specific PCR: a sensitive technique for accurate evaluation of CpG methylation.. https://doi.org/10.1016/j.bbrc.2003.08.005

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