Methylated DNA occurs naturally in prokaryotes and eukaryotes. In humans, it has been implicated in epigenetics, genomic imprinting, embryonic development, X-chromosome inactivation and carcinogenesis. In humans, a majority of all cytosines in the dinucleotide 5’-CpG-3’ are methylated, though non-CpG methylation patterns have also been found. A breakthrough in the detection of DNA methylation was made with the discovery that bisulfite has higher specificity for cytosine than 5-methyl cytosine. The DNA after conversion is single stranded and non-complementary.
Intended Use:This Genomic DNA Methylation Detection BioAssay™ Kit is developed and optimized for sodium bisulfite treatment.
Range:2ug-500pg
Test Principle:This DNA methylation detection kit offers a fast, efficient and convenient alternative to traditionalbisulfite conversions. This kit provides users with a true endogenous human positive control which eliminates any ambiguity in validating results. The positive control has been tested in over 15 different human tissues . The protocol is optimized to minimize DNA degradation, reagent preparation, and conversion time. The protocol guarantees ~99% conversion of cytosine to uracil and ~99% CpG protection, as calculated by sequencing, and is sensitive over a range of 2ug-500pg of DNA. In <3 hours, users can obtain high quality bisulfite modified DNA ready for analysis by PCR, sequencing, or microarrays. The kit is enough for 50 reactions.
Kit Components:Conversion Reagent (CVR)*, 5x0.4g, RTActivator Solution (ACS)*, 1x5ml, RTStabilizer Solution (STS)*, 1x500ul, RTEquilibration Buffer (EQB), 1x20ml, RTDesulfonation Buffer (DSB), 1x4ml, RT Elution Buffer (TE), 2x1ml, RT Control MetPrimers (CMP), 5uM, 1x100ul, -20ºCControl Human DNA, 2ug, 1x100ul, -20ºCColumn 50x, RT*After mixing ACS, STS, and CVR to prepare Conversion Buffer (CVB), store at -20ºC.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
