Patent ID: 7485443

Claim:
A method for detecting polymorphisms in unamplified DNA, comprising: a) providing: i) unamplified DNA, wherein said unamplified DNA comprises 50 micrograms or less of genomic DNA wherein said genomic DNA has not been subjected to an in vitro amplification method; ii) reagents for conducting an invasive cleavage reaction, wherein said reagents comprise a plurality of oligonucleotide pairs, wherein each of said oligonucleotide pairs comprises a probe oligonucleotide and an upstream oligonucleotide, wherein said probe oligonucleotide associates and dissociates from a target sequence, wherein said upstream oligonucleotide is configured to hybridize to a target sequence in a manner that overlaps with said probe oligonucleotide, wherein said probe and upstream oligonucleotides are attached to the same solid surface, wherein said upstream oligonucleotide is attached to said surface via a spacer molecule that has a length of 10 hexaethylene molecules, and wherein the ratio of probe oligonucleotide to upstream oligonucleotide is from 5:1 to 30:1; and b) treating said unamplified DNA with said reagents under conditions such that invasive cleavage reactions occur on said solid surface with said unamplified DNA and said reagents such that the presence or absence of a plurality of different polymorphisms in said genomic DNA are identified, and wherein said treating step is carried out in a single reaction vessel.