Patent ID: 6235502
Filing Date: 2001-05-22
Classification: C12Q,Y10T

Abstract:
A method for isolating mismatched DNA containing fragments nicked by MutSLH from two DNA samples comprising:(a) digesting a first DNA sample with a restriction enzyme to obtain DNA fragments;(b) ligating Y-shaped adapters to the fragments to obtain fragment-plus-adapter constructs;(c) repeating steps (a) through (b) with a second sample;(d) methylating the products formed from step (b) of the first sample but not the second;(e) mixing the methylated and unmethylated products obtained from the samples, denaturing, and reannealing to form hemimethylated heterohybrids;(f) treating the fragments produced in step (e) with a MutSLH preparation in order to nick DNA containing mismatches and form a 3'-OH end; and(g) elongating the 3'-OH end with one or more DNA polymerase;(h) isolating the elongated, nicked DNA to provide isolated mismatched DNA.