Patent ID: 7947446
Filing Date: 2011-05-24
Classification: C12Q

Abstract:
1. A method of simultaneously screening for genetic mutations in different genes in a multitude of samples, the method comprising: providing a sample of heteroduplex molecules, wherein the heteroduplex molecules each include: wherein the sample forms: incubating the sample of heteroduplex molecules with a ribonuclease enzyme, wherein the ribonuclease enzyme cleaves the unhybridized ribonucleotides in the second population and the third population and exposes a 3′ hydroxyl group at the cleavage sites; protecting the 3′ end of the RNA strand; performing differential sequence fill-in at the cleaved sites using an RNA-primed DNA polymerase to create a second sample; and incubating the second sample with a single-strand-specific nuclease to digest the unprotected deoxyribonucleotides in the third population and in the second population not filled in with a complementary dNTP; wherein the first population and heteroduplexes in the second population filled in with a complementary dNTP are undigested; incubating the undigested portion of the second sample with: wherein the DNA-dependent DNA polymerase creates: linking a tagged reporter adapter to the unprotected overhang to form marked mutant-adapter hybrids; and detecting said marked mutant-adapter hybrids.