Methods of detecting mutations in BRAF and epigenetic changes

摘要

Disclosed are methods for assessing the methylation and mutation status of nucleic acid in a sample. The methods provide for methylation-dependent modification of the nucleic acid in a sample, and subsequently nucleic acid amplification processes to distinguish between mutated and non-mutated target sequence.

主权项

1. A method for detecting a mutated BRAF target sequence of interest in a DNA-containing sample, the method comprising:
(a) contacting the DNA-containing sample with a modifying reagent which selectively modifies non-methylated cytosine residues in the DNA to produce detectable modified residues but which does not modify methylated cytosine residues, (b) amplifying the modified DNA-containing sample to produce an amplified target sequence using at least one primer that hybridizes specifically to the mutated BRAF target sequence in the modified DNA-containing sample, and (c) detecting the amplified target sequence, thereby detecting the mutated BRAF target sequence.