US 2003/0190609 A1 · Address/capture tags for flow-cytometery based minisequencing

Abstract

A method for generating address/capture tags for use in a sensitive and rapid flow-cytometry based assay for the multiplexed analysis of SNPs based on polymerase-mediated primer extension using microspheres as solid supports is described. Single-nucleotide polymorphisms (SNPs) are the most abundant type of human genetic variation. These variable sites are present at high density in the genome, making them powerful tools for mapping and diagnosing disease-related alleles. Subnanomolar concentrations of sample in small volumes (10 ml) can be analyzed at rates greater than one sample per minute, without a wash step. Genomic analysis using multiplexing microsphere arrays, enables the simultaneous analysis of dozens, and potentially hundreds of SNPs per sample. The method has been tested by genotyping the Glu69 variant from the HLA DPB1 locus, a SNP associated with chronic beryllium disease, as well as HLA DPA1 alleles.

Claims13

Dependent claims

A method for identifying a set of sequences useful as address/capture tags which comprises the steps of:

(a) generating a chosen number of single-stranded, random oligonucleotide sequences having a chosen length;
(b) rejecting all sequences from said chosen number of single-stranded, random oligonucleotide sequences having common subsequences with a subsequence length greater than a chosen number of bases, the remaining sequences forming a first group of sequences;
(c) rejecting all sequences in said first group of sequences which can form stable hairpins, the remaining sequences forming a second group of sequences;
and
(d) rejecting all sequences in said second group of sequences which can form stable dimers, the remaining sequences forming a third group of sequences;
whereby a set of sequences is identified such that the sequences, if synthesized, would hybridize to their respective complements with a high degree of specificity.

3 more elements

Dependencies are inferred from the claim text.

Address/capture tags for flow-cytometery based minisequencing

Life of this patent

Address/capture tags for flow-cytometery based minisequencing

Life of this patent

Abstract

Claims13