Method for nucleic acid isolation and amplification
First Claim
Patent Images
1. A method for amplifying a polynucleotide molecule of interest or a fragment thereof, comprising:
- (a) isolating a polynucleotide molecule from a nucleic acid population using an immobilizable separation group to provide an isolated polynucleotide molecule, and (b) isothermally amplifying the isolated polynucleotide molecule or a fragment thereof.
3 Assignments
0 Petitions
Accused Products
Abstract
The present invention provides methods and compositions for sequence-specific isolation of polynucleotide molecules from nucleic acid populations and subsequent amplification of isolated polynucleotide molecules or fragments thereof.
142 Citations
55 Claims
-
1. A method for amplifying a polynucleotide molecule of interest or a fragment thereof, comprising:
-
(a) isolating a polynucleotide molecule from a nucleic acid population using an immobilizable separation group to provide an isolated polynucleotide molecule, and (b) isothermally amplifying the isolated polynucleotide molecule or a fragment thereof. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44)
-
-
45. A method for amplifying multiple polynucleotide molecules of interest from a population of nucleic acid molecules, comprising:
-
(a) isolating multiple polynucleotide molecules from a nucleic acid population using one or more immobilizable separation groups to provide isolated polynucleotide molecules of interest, and (b) isothermally amplifying the isolated polynucleotide molecules or fragments thereof. - View Dependent Claims (46, 47, 48, 49, 50, 51, 52)
-
-
53. A method for amplifying a genomic DNA molecule of interest or a fragment thereof, wherein the genomic DNA molecule of interest comprises a polymorphic sequence, comprising:
-
(a) contacting a genomic DNA population that comprises the genomic DNA molecule of interest with an oligonucleotide, wherein (i) the oligonucleotide comprises a sequence at least substantially complementary to a target nucleic acid sequence in one strand of the genomic DNA molecule of interest, (ii) the target nucleic acid sequence is located immediately 3′
to the polymorphic sequence in the one strand of the genomic DNA molecule of interest, and(iii) the 3′
portion of the oligonucleotide is complementary to the polymorphic sequence or a portion thereof when annealing to the one strand of the genomic DNA molecule of interest,(b) extending the oligonucleotide in the presence of an immobilizable nucleotide using the one strand of the genomic DNA molecule of interest to which the oligonucleotide anneals as a template to provide an extension product, (c) immobilizing to a substrate via the immobilizable nucleotide the extension product to which the genomic DNA molecule of interest is bound, (d) removing the immobilized extension product to which the genomic DNA molecule of interest is bound to thereby isolate the genomic DNA molecule of interest from the genomic DNA population, (e) optionally elute the genomic DNA molecule of interest from the substrate, and (f) isothermally amplifying the isolated or eluted genomic DNA molecule of interest or a fragment thereof.
-
-
54. A method for assembling a haplotype comprising:
-
(a) providing a nucleic acid population from an organism for which a haplotype is of interest;
(b) separately isolating polynucleotide molecules by haplotype-specific extraction using multiple substrates, wherein (i) polynucleotide molecules are isolated at multiple extraction sites using each substrate, (ii) no polynucleotide molecules isolated at one extraction site using one substrate comprise a polymorphic site also present in polynucleotide molecules isolated at other extraction sites using the same substrate, (iii) one or more polynucleotide molecules isolated at one extraction site using one substrate comprise a polymorphic site also present in polynucleotide molecules isolated at a neighboring extraction site using other substrates, (c) separately characterizing polymorphic sites in polynucleotide molecules isolated using each substrate;
(d) assembling a haplotype based on the characterization of polymorphic sites present in polynucleotide molecules isolated using more than one substrate. - View Dependent Claims (55)
-
Specification