EpiTect MethyLight PCR +ROX Kit (200)

Designed by QIAGEN for quantitative, real-time probe-based PCR analysis of DNA methylation status, this kit is supplied in a convenient master mix format with ROX passive reference dye included directly in the mix. It is intended for real-time cyclers that require a high concentration of ROX dye for fluorescence normalization, such as instruments from Applied Biosystems (excluding the Applied Biosystems 7500 Real-Time PCR Systems).

The chemistry is specifically developed for the demands of bisulfite converted DNA. It uses sequence-specific probes — including TaqMan-style dual-labeled probes with a 5' fluorophore and an internal or 3' quencher — and supports both single and duplex reactions. The master mix buffer contains a specially optimized combination of KCl and (NH4)2SO4, along with Factor MP and balanced cations, which promote a high ratio of specific-to-nonspecific primer and probe binding during annealing. This enables discriminative hybridization and reliable probe-based detection of methylated and unmethylated target sequences in bisulfite converted DNA.

The kit can be used in combination with EpiTect MethyLight Assays or other dual-labeled assays for both quantitative and semiquantitative methylation analysis, and is suitable for most real-time thermal cyclers other than the Applied Biosystems 7500. Validated protocols are available to help ensure optimal results across supported instruments.