Bisulfite conversion of DNA
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.
1. A method for bisulfite conversion of DNA, comprising reacting genomic DNA with a bisulfite reagent, wherein the reaction is carried out in the presence of a compound having the following formula:
2. The method of claim 1 , wherein the compound comprises an n-alkylene glycol.
3. The method of claim 2 , wherein the compound comprises a dialkyl ether.
4. The method of claim 3 , wherein the compound comprises diethylene glycol dimethyl ether (DME).
5. The method of claim 1 , wherein the compound is present in a concentration of about 1-35%.
6. The method of claim 1 , wherein the compound is present in a concentration of about 5-25%.
7. The method of claim 1 , further comprising analysis of the converted DNA, wherein analysis comprises a polymerase chain reaction.
8. The method of claim 1 , wherein the DNA is extracted from a of tissue sample or bodily fluid.
9. The method of claim 1 , further comprising diagnosis or prognosis of an adverse event in a subject in need thereof, wherein the adverse event is selected from the group consisting of: drug interactions; cancer diseases; CNS malfunctions, damage or disease; dementia; cardiovascular disease, malfunction and damage;
malfunction, damage or disease of the gastrointestinal tract; malfunction, damage or disease of the respiratory system; lesion, inflammation, infection, or immunity; malfunction, damage or disease of the body as an abnormality in the development process; malfunction, damage or disease of the skin, of the muscles, of the connective tissue or of the bones; endocrine and metabolic malfunction, damage or disease; headaches or sexual malfunction.
10. The method of claim 1 , further comprising distinguishing cell or tissue type, or analyzing cell differentiation.