MethylEasy™ Xceed (ME002)

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

Genomic DNA was isolated using the Genomic DNA Isolation Kit (Qiagen, Valencia, CA) from UMR-106 cells incubated with 1,25(OH)2D3 or vehicle. About 1 µg of rat genomic DNA was treated with bisulfite conversion reagents using the MethylEasy Xceed Rapid DNA Bisulfite Modification Kit (Human Genetic Signatures, Australia) according to the manufacturer’s instruction. The bisulfite converted genomic DNA was amplified using BMP2-F1 (5′-AGTGGGGGAATTTAGAGGTAAG-3′) and BMP2-R1 (5′-TATCRTACCRACCCCTCRAAACCCC-3′) primers with the following cycling condition: 95°C 3 min, then 30 cycles of 95°C 30 sec, 56°C 30 sec and 72°C 60 sec. About 2% of the PCR products were subject to a second round of amplification using nested primers BMP2-F2 (5′-AGAGGTAAGTTTATTTTGAATTTG-3′) and BMP2-R2 (5′-CTTCRTCCCRAACTACCAATCCC-3′) with the same PCR procedural condition as above. The amplified PCR products were then subcloned; 25 clones from each ligation were randomly picked and sequenced on an Applied Biosystems 3130 instrument. Fractional DNA methylation [#C/(#C+#T)] for each CpG site was determined based on the number of times it was sequenced as a T or as a C.

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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 - UMR-106 BMP2 using MethylEasy™ Xceed (ME002) from Genetic signatures.

Manufacturer protocol

Download the product protocol from Genetic signatures for MethylEasy™ Xceed (ME002) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - UMR-106 BMP2 using MethylEasy™ Xceed (ME002) from Genetic signatures. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.