PyroMark Q24 CpG MLH1 (4 x 24)

Designed by QIAGEN, this sequence-based Pyrosequencing assay enables quantification of methylation at CpG sites within the MLH1 gene. The kit provides forward and reverse PCR primers together with a sequencing primer for amplification and analysis of bisulfite-converted DNA. The reverse primer is biotinylated, allowing isolation of the correct template for the sequencing reaction, and the sequencing primer additionally serves as a control for complete bisulfite conversion. The target region is located within chr3:36,993,279-36,993,309 (reference genome GRCh38.p14) of the MLH1 gene (Ensembl ID: ENSG00000076242).

The workflow comprises four steps: bisulfite conversion of sample DNA, PCR amplification, preparation of a single-stranded DNA template, and Pyrosequencing analysis on a compatible instrument. The assay is intended for use with the PyroMark Q24, PyroMark Q24 Advanced, and PyroMark Q48 Autoprep instruments. Each PCR primer is dissolved in 120 µL of high-purity water to give a 10 µM concentration, and each sequencing primer vial is dissolved in 180 µL of buffer to give a 10 µM concentration. The kit is shipped on dry ice and should be stored at −20 °C upon arrival.

The product is intended for molecular biology applications and is not intended for the diagnosis, prevention, or treatment of a disease.