The Quest 5-hmC Detection Kit™ from Zymo Research allows for sequence specific detection of 5-hydroxymethylcytosine within DNA using a simple and efficient reaction setup. Utilizing a robust and highly specific 5-hmC Glucosyltransferase enzyme, 5-hydroxymethylcytosine in DNA is specifically tagged with a glucose moiety yielding a modified base, glucosyl-5-hydroxymethylcytosine.
- Method to specifically distinguish 5-hydroxymethylcytosine in sequence and locus specific context.
- Adaptable for use with any glucosyl-5-hydroxymethylcytosine sensitive endonucleases (e.g. GlaI, MspI, TaqαI, McrBC)
- Convenient and reliable one tube reaction.
- Processed DNA eluted in water or low salt buffer is suitable for analysis by a variety of downstream molecular biology applications.
Treatment of DNA containing 5hmC with 5-hmC Glucosyltransferase specifically adds a glucose moiety yielding glucosyl-5-hydroxymethylcytosine. Subsequent digestion with glucosyl-5-hydroxymethylcytosine sensitive restriction endonucleases will cut DNA with 5-methylcytosine or 5-hydroxymethylcytosine in their recognition sequence, but leave glucosyl-5-hydroxymethylcytosine DNA uncleaved.
After glucosylation of 5-hydroxymethylcytosine, digestion of DNA with “glucosyl-5-hydroxymethylcytosine sensitive” restriction endonucleases, or GSRE’s, allows for effective differentiation of 5-methylcytosine from 5-hydroxymethylcytosine. Identification of 5-hydroxymethylcytosine in a sequence specific context can then be deduced from the restriction endonuclease recognition sequence.View Quest 5-hmC Detection Kit™ on Zymo Research’s website