EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification 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

Methylation-specific polymerase chain reaction (PCR)
Total cell DNA was isolated from the cultured cells using a QIAamp DNA Mini kit (Qiagen N.V., Venlo, The Netherlands). The genomic DNA was modified as described previously () using a DNA bisulfite modification kit (Chemicon, Temecula, CA, USA). The methylated CpG dinucleotides within a CpG island in the promoter region of FHIT were detected using the following specific PCR primer sets: methylated FHIT forward, 5-TTGGGGCGCGGGTTTGGGTTTTTACGC-3 and reverse, 5-CGTAAACGACGCCGACCCCACTA-3; and unmethylated FHIT forward, 5-TTGGGGTGTGGGTTT GGGTTTTTATG-3 and reverse, 5-CATAAACAACACCAACCCCACTA-3 (). Each PCR mixture system contained 10X PCR buffer (Qiagen N.V.), deoxynucleotide triphosphates (1.25 mmol/l), primers (0.6 mmol/l), 1 unit HotStarTaq (Qiagen N.V.), and bisulfite-modified DNA (100 ng). PCR was performed in a programmable thermal controller (MJ Research, Inc., Quebec, Canada) under the following conditions: Preheating at 94°C for 3 min, 94°C for 30 sec, 62°C for 30 sec and 72°C for 30 sec for 38 cycles and a final extension at 72°C for 7 min. The PCR products were analyzed on a 2% agarose gel.

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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 - HepG2 FHIT using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit from Fisher Scientific.

Manufacturer protocol

Download the product protocol from Fisher Scientific for EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - HepG2 FHIT using EMD Millipore™ Chemicon™ CpGenome™ Universal DNA Modification Kit from Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.