Method for analysis of oligonucleotide analogs
First Claim
1. A method of separating a population of duplexes, each comprising one of a population of different, substantially uncharged oligomeric analyte molecules and a specific probe molecule,wherein said substantially uncharged analyte molecules are oligonucleotide analogs composed of linked subunits of which at least 90% are uncharged, and said specific probe molecule is a fully charged nucleic acid or fully charged nucleic acid analog,the method comprising:
- (a) applying to an ion exchange medium a mixture of (i) the different substantially uncharged analyte molecules and (ii) the specific probe molecule, under conditions such that the probe forms stable duplexes with a plurality of or all of the different substantially uncharged analyte molecules,thereby forming a plurality of different probe-analyte duplexes, which differ from each other with respect to the presence, length or position of an unhybridized portion of the probe molecule, and(b) separating said different probe-analyte duplexes from each other and from single stranded analyte or probe molecules within the medium.
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Abstract
Disclosed are methods for separating, detecting, quantitating and/or isolating predominantly uncharged oligonucleotide analogs, by resolving duplexes of such molecules with complementary or near-complementary DNA or charged DNA analogs. The DNA or charged analog may be labeled for detection purposes.
13 Citations
19 Claims
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1. A method of separating a population of duplexes, each comprising one of a population of different, substantially uncharged oligomeric analyte molecules and a specific probe molecule,
wherein said substantially uncharged analyte molecules are oligonucleotide analogs composed of linked subunits of which at least 90% are uncharged, and said specific probe molecule is a fully charged nucleic acid or fully charged nucleic acid analog, the method comprising: -
(a) applying to an ion exchange medium a mixture of (i) the different substantially uncharged analyte molecules and (ii) the specific probe molecule, under conditions such that the probe forms stable duplexes with a plurality of or all of the different substantially uncharged analyte molecules, thereby forming a plurality of different probe-analyte duplexes, which differ from each other with respect to the presence, length or position of an unhybridized portion of the probe molecule, and (b) separating said different probe-analyte duplexes from each other and from single stranded analyte or probe molecules within the medium. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification