OneStep PLUS qMethyl PCR Kit

No bisulfite conversion needed: from DNA sample to results in just one step.Accurate quantification of DNA methylation levels at the genomic regions of choice.Ideal for rapid screening of DNA methylation at single and multi-locus.The OneStep PLUS qMethyl PCR Kit provides a simple, straightforward, and bisulfite-free procedure for the quantification of DNA methylation at custom-selected genomic regions. This bisulfite-free assessment is accomplished by splitting the DNA to be tested into two parts: a Test Reaction and a Reference Reaction. DNA in the Test Reaction is digested with Methylation Sensitive Restriction Enzymes MSREs while DNA in the Reference Reaction is not. The DNA from both reactions is then amplified using Real-Time PCR. The difference in Cycle Threshold Ct values between the Reference and Test reactions is used to calculate the percentage of DNA methylation at the selected genomic region. Compatible Thermal Cyclers: Bio-rad CFX-96, CFX Opus 96, QuantStudio qPCR Systems, Roche LightCycler 480, or similar.Detection Dye: Provided dye is compatible with SYBR Green Filter. Assay is flexible with other dyes and TaqMan probes.Input: High quality DNA, suitable for use in restriction enzyme digestion.A260/280 > 1.8, A260/230 > 1.8.Optimal DNA input is 20 ng in 5 l.Range validated 5 ng 1 g in 5 l.Processing Time: Optimal 14 hours.Storage: Store at -20C for up to 12 months. Avoid repeated freeze-thaw cycles of reagents. Prolong storage at -80C.2X Test PreMix and 2X Reference PreMix are light sensitive. Minimize light exposure. Shipping Weight: 0.0399 lbsShipping Dimensions: 5.00 x 0.50 x 6.00Research or further manufacturing use only, not for food or drug use.