Method of rapidly quantifying global DNA methylation

Abstract

This invention is related to a method and assay kit for rapidly quantifying global DNA methylation through immobilizing DNA by simple dry-capture on the plastic carrier followed by immunodetection of 5-methylcytosine structure that is the marker of DNA methylation.

1 . A method of rapidly quantifying global DNA methylation comprising:
 a) immobilization of DNA with a binding buffer onto a plastic carrier coated with poly-L-lysine at concentration of 0.001-0.1% by two-phase temperature for said an appropriate period.   b) immunodetection of 5-methylcytosine structure contained in the DNA.   
     
     
         2 - 8 . (canceled) 
     
     
         9 . The method according to  claim 1  wherein said buffer is tris-EDTA solution. 
     
     
         10 . The method according to  claim 1  wherein said two-phase temperature is consisting of first phase at 25-42° C. followed by second phase at 50-75° C. 
     
     
         11 . The method according to  claim 1  wherein said two-phase temperature is consisting of first phase at 37° C. followed by second phase at 60° C. 
     
     
         12 . The method according to  claim 1  wherein said an appropriate period is from 2 to 4 hours. 
     
     
         13 - 19 . (canceled) 
     
     
         20 . The method according to  claim 1  wherein said the kit is comprised of:
 a) the microwell plate or strips coated with poly-L-lysine.   b) the concentrated washing buffer comprising of phosphate salt and 0.1% tween-20.   c) the DNA immobilization buffer comprising of Tris-HCL and EDTA.   d) the blocking buffer comprising of phosphate salt and 2% BSA   e) the color development solution comprising of TMB and H 2 O 2      f) the stop solution comprising of HCl   g) a positive control from the enzymatically methylated genomic DNA or naturally methylated virus DNA   h) instructions for conducting an assay according to the method of this invention