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.

Protocol tips
For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation.

Publication protocol

Quantitative methylation-specific PCR (qMSP). Bisulfite
treatment of genomic DNA was performed using EpiTect
Bisulfite Kit (Qiagen). Quantitative real-time MSP was carried
out with bisulfite-modified genomic DNA as a template using
specific primer sequences for the methylated or unmethylated
forms of each gene. The following methylated/unmethylatedspecific primers were used: 5'-GAGTCGGCGGTTAGGA
TGC-3' (methylated forward) and 5'-GAGTCGGCGGTTA
GGATGT-3' (unmethylated forward) and 5'-TTTAAAAAC
AAAAAACTCTTTAACCCAAA-3' (reverse). For qMSP,
20 µl of reaction mixture containing 2 µl (10-100 ng/µl) of
bisulfite-treated DNA, 10 µl of SYBR Premix EX Taq (Takara
Bio), 0.4 µl of Rox reference dye (50X), and 200 nM of each
primer was reacted using an 7500 Fast Real-time PCR system
at 95˚C for 30 sec, and then repeated for 40 cycles (at 95˚C for
3 sec, and at 62˚C for 30 sec) for amplification. The PCR
product was reacted at 95˚C for 15 sec, at 60˚C for 1 min, and
at 95˚C for 15 sec to examine specificity. Methylation and
unmethylation of the specific CpG site were calculated as
follows (Ct represents the threshold cycle):
Percentage of methylation = 100/(1 + 2ΔCt methylation - ΔCt unmethylation) %.

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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 - SKOV3 AGR2 using EpiTect Bisulfite Kit from Qiagen.

Manufacturer protocol

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

Download manufacturer protocol

Videos

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