Patent ID: 7303869

Claim:
A method for detecting polymorphisms in unamplified DNA, comprising: a. providing: i. unamplified genomic DNA, wherein said unamplified genomic DNA comprises 50 micrograms or less of genomic DNA; ii. a microsphere; iii. oligonucleotides and enzymes for conducting an invasive cleavage reaction, wherein said oligonucleotides comprise a probe oligonucleotide and an upstream oligonucleotide each attached to said microsphere via a spacer molecule, wherein said spacer molecule has a length of 10 hexaethylene glycol molecules and, wherein said upstream oligonucleotide is configured to hybridize to a target sequence in a manner that overlaps with said probe oligonucleotide; and b. treating said unamplified genomic DNA with said oligonucleotides and enzymes under conditions such that said upstream oligonucleotide and said probe oligonucleotide hybridize to said unamplified genomic DNA such that said upstream oligonucleotide overlaps said probe oligonucleotide, and an invasive cleavage reaction occurs on said microsphere such that the presence or absence of a polymorphism in said unamplified genomic DNA is identified.