EpiScope MSP 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

Quantitative RT-PCR, methylation-specific PCR, and bisulfide sequencing PCR
Quantitative RT-PCR (qPCR) was carried out with first-strand cDNAs of the cell lines and SYBR PremixEx TaqII (Takara Bio, Otsu, Japan) using the Viia7 system (Life Technologies, Carlsbad, CA, USA). Unmethylated cytosines in the genomic DNAs from the cell lines were converted to uracil by bisulfide reactions using the EZ DNA Methylation-Gold Kit (ZYMO Research, Irvine, CA, USA). Methylation-specific PCR (MSP) was carried out using the TaKaRa EpiScope MSP Kit. Bisulfide sequencing PCR (BSP) was carried out using the bisulfide-treated genomic DNAs and EpiTaq HS (Takara). The PCR products were inserted into the pGEM-T easy vector (Promega KK, Tokyo, Japan) and transformed into DH5α, and plasmids isolated from 10 colonies of each sample were sequenced.

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Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA methylation profiling Gene specific profiling - HEK 293T NELL1 using EpiScope MSP Kit from Takara Bio Inc.

Manufacturer protocol

Download the product protocol from Takara Bio Inc for EpiScope MSP Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - HEK 293T NELL1 using EpiScope MSP Kit from Takara Bio Inc. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.