Book Chapter

Closed-Tube PCR Methods for Locus-Specific DNA Methylation Analysis

Ida LM Candiloro, Thomas Mikeska, Alexander Dobrovic

EPIGENETICS PROTOCOLS, SECOND EDITION | Methods in Molecular Biology | HUMANA PRESS INC | Published : 2011


Closed-tube PCR methods (sometimes referred to as in-tube PCR methods) for locus-specific DNA -methylation analysis are methodologies in which the amplification and analysis of bisulphite-modified DNA take place in one tube without the need to remove the PCR products for further analysis. Closed-tube methodologies lend themselves to high-throughput applications and molecular diagnostics but are also applicable as a research tool. We review three closed-tube methodologies, methylation-sensitive high-resolution melting (MS-HRM), MethyLight, and sensitive melting after real-time analysis - methylation-specific PCR (SMART-MSP). Closed-tube detection can be performed by simultaneously amplifying ..

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University of Melbourne Researchers