Bisulfite conversion of DNA
OwnerKurt Berlin, Matthias Ballhause +1inventors
InventorsKurt Berlin, Matthias Ballhause, Karen Cardon
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Abstract
The present invention provides an improved method for the bisulfite conversion of DNA, and facilitates the analysis of cytosine methylation of genomic DNA. Novel combinations of denaturing solvents, new reaction conditions and new purification methods provide surprisingly efficacious methods for bisulfite conversion of DNA relative to prior art methods. The converted DNA may subsequently be analyzed by many different methods.
Claims21
A method for bisulfite conversion of DNA, comprising reacting genomic DNA with a bisulfite reagent, wherein said reaction is carried out in the presence of a compound selected from the group consisting of dioxane, one of its derivatives, and a similar aliphatic cyclic ether.
A method for bisulfite conversion of DNA, comprising reacting genomic DNA with a bisulfite reagent, wherein said reaction is carried out in the presence of a compound with the following formula:
- n=1-35000
- m=1-3
- R1=H, Me, Et, Pr, Bu
- R2=H, Me, Et, Pr, Bu
A method for bisulfite conversion of DNA, comprising reacting isolated genomic DNA with a bisulfite reagent, wherein the reaction is conducted at a temperature in the range of 0-80° C., and wherein the reaction temperature is briefly increased to a range of about 85-100° C. during the course of the conversion.
A method for the bisulfite conversion of DNA, comprising reacting DNA with a bisulfite reagent, and purifying the converted DNA by means of ultrafiltration.
A kit, comprising:
- a reagent containing bisulfite;
- denaturing reagents or solvents, a radical scavenger, and primers for the production of amplificates.
Dependencies are inferred from the claim text.