Methods of identification and isolation of polynucleotides containing nucleic acid differences
AssigneeGensat S.A.
InventorGilbert Thill
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Abstract
The present invention is directed to methods of isolation of related polynucleotides harboring nucleic acid difference within a polynucleotide sample. The method will be useful in detecting and identifying alternative splicing events and corresponding splicing isoforms and to detect genomic DNA differences between genomes. The method according to the present invention is based on the use of a single-stranded trap. The single-stranded trap preferably involves the use of single-strand binding protein.
Claims23
A method of isolating or selecting related DNA molecules harboring nucleic acid differences in a DNA sample, said method comprising the selection of heteroduplexes containing at least one internal single stranded region with a single stranded trap, wherein
- i) said heteroduplexes are formed between said related DNA molecules,
- ii) said internal single stranded regions represent said nucleic acid differences, and
- iii) the single stranded trap involves the use of a Recognition Element having preferential affinity for single-stranded DNA compared to double stranded DNA that is a single strand binding protein.
A method of isolating or selecting related DNA harboring nucleic acid differences in a DNA sample, comprising the following steps:
- (a) obtaining a DNA sample containing said related DNA molecules;
- (b) denaturing DNA molecules present in said sample;
- (c) annealing said denatured DNA molecules to allow the formation of heteroduplexes between said related DNA molecules harboring nucleic acid differences;
- and,
- (d) isolating or selecting heteroduplexes containing at least one internal single stranded region using a single-stranded trap comprising a Recognition Element that is a single strand DNA binding protein.
Dependencies are inferred from the claim text.