METHODS FOR ESTIMATING THE SIZE OF DISEASE-ASSOCIATED POLYNUCLEOTIDE REPEAT EXPANSIONS IN GENES

摘要

Methods for estimating the size of disease-associated polynucleotide repeat expansions in genes are disclosed which use restriction enzymes that do not cut within a repeat expansion and which are frequent cutting restriction enzymes that cut genomic DNA outside of the expansion into fragments of a size below the threshold capable of detection. A hybridisation probe that can bind to multiple sites within the expansion is then used to estimate its length and to correlate that to the diagnosis or prognosis of disease.

主权项

1. A method of estimating the size of a disease-associated polynucleotide repeat expansion in a gene, the method comprising:
(a) contacting the sample of genomic DNA from an individual with one or more restriction enzymes, wherein the restriction enzymes have restriction sites flanking the region of genomic DNA containing the polynucleotide repeat expansion and are capable of cutting the genomic DNA outside of the fragment containing the polynucleotide repeat expansion into a plurality of DNA fragments; (b) optionally separating the nucleic acid fragment containing the polynucleotide repeat expansion from the plurality of DNA fragments; (c) contacting the nucleic acid fragment containing the polynucleotide repeat expansion with a hybridisation probe capable of targeting multiple sites within the polynucleotide repeat expansion; and (d) detecting the hybridisation of the hybridisation probe to the polynucleotide repeat expansion to estimate the size of the disease-associated polynucleotide repeat expansion; wherein the one or more restriction enzymes do not cut within the repeat expansion and are frequent cutting restriction enzymes capable of cutting genomic DNA into fragments of a modal size below the size of the repeat expansion, and wherein the disease associated with the polynucleotide repeat expansion is a neurological disease.