EpiTect Bisulfite Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation.	Always include fully methylated (in-vitro methylated DNA) and non-methylated controls (Leucocyte DNA, DNMT1/3 double KO DNA) during MSP

Upstream tips
For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation.
Protocol tips
Always include fully methylated (in-vitro methylated DNA) and non-methylated controls (Leucocyte DNA, DNMT1/3 double KO DNA) during MSP

Publication protocol

MSP assay
Genomic DNA was extracted from samples with a DNeasy tissue kit (Qiagen). Bisulfite reaction was performed using 5 μg of genomic DNA with the EpiTect Bisulfite Kit (Qiagen) according to manufacturer’s instruction. Then MSP was performed following the methods introduced in one previous study [] with methylated-specific primers (M): forward, 5’-TTTTCGTTTTCGTTATTAAGGTTTC-3’ and reverse, 5’-GACCCAACCTAACTTAACTTCGTC-3’ (product size: 239bp); and unmethylated-specific primers (U): forward, 5’-TTTTTGTTTTTGTTATTAAGGTTTT-3’ and reverse, 5’- CAACCCAACCTAACTTAACTTCATC-3’ (product size: 240bp). PCR products were verified by 2.5% agarose gel electrophoresis.

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Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA methylation profiling Gene specific profiling - SiHa miR-375 using EpiTect Bisulfite Kit from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for EpiTect Bisulfite Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - SiHa miR-375 using EpiTect Bisulfite Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.