Individually synthesized primers to be used in whole genome amplification
AssigneeQiagen Gaithersburg, Inc.
InventorsIrina Nazarenko, Dominic O'Neil, Ha Thai, Attila Lorincz
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Abstract
Disclosed are compositions and methods for random amplification of nucleic acid sequences of interest using random-G-deficient primers. Also disclosed are methods of randomly amplifying a target nucleic acid sequence using random G-deficient primers alone or in combination with random, partially random, or specific primers.
Claims27
A method of randomly amplifying a target nucleic acid sequence, the method comprising, bringing into contact a set of primers, DNA polymerase, and a target sample, wherein the primers are random G-deficient primers, and incubating the target sample under conditions that promote replication of the target sequence, wherein replication of the target sequence results in replicated strands.
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A method of randomly amplifying a target nucleic acid sequence, the method comprising, bringing into contact a set of primers, DNA polymerase, and a target sample, wherein the primers are random G-deficient primers, and incubating the target sample under conditions that promote replication of the target sequence, wherein nucleic acids in the target sample are not separated from other material in the target sample.
A method of randomly amplifying messenger RNA, the method comprising,
- reverse transcribing messenger RNA to produce a first strand cDNA,
- bringing into contact a set of random G-deficient primers, DNA polymerase, and the first strand cDNA, and incubating under conditions that promote replication of the first strand cDNA,
- wherein replication of the first strand cDNA results in replicated strands, wherein during replication at least one of the replicated strands is displaced from the first strand cDNA by strand displacement replication of another replicated strand.
A method of randomly amplifying a target nucleic acid sequence, the method comprising,
- (a) mixing a set of random G-deficient primers with a target sample, to produce a primer-target sample mixture, and incubating the primer-target sample mixture under conditions that promote hybridization between the random G-deficient primers and the target sequence in the primer-target sample mixture,
- (b) mixing DNA polymerase with the primer-target sample mixture, to produce a polymerase-target sample mixture, and incubating the polymerase-target sample mixture under conditions that promote replication of the target sequence,
- wherein replication of the target sequence results in replicated strands, wherein during replication at least one of the replicated strands is displaced from the target sequence by strand displacement replication of another replicated strand,
- wherein the target sequence is a nucleic acid sample of substantial complexity.
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Dependencies are inferred from the claim text.