Multiplex amplification methods
First Claim
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1. A method for amplifying an RNA target comprising:
- (a) obtaining a capture probe comprising from the 5′
end a second priming sequence, a tag sequence, a first priming sequence and a sequence complementary to the RNA target;
(b) hybridizing the RNA target to the capture probe and extending the RNA using a polymerase and using the capture probe as template to form a duplex between the capture probe and the extended RNA target;
(c) optionally destroying or removing the capture probe; and
(d) amplifying the portion of the extended RNA comprising the first and second primer regions and the tag region.
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Abstract
Compositions and methods for amplifying selected polynucleotides, including DNA and RNA, particularly in multiplex amplification reactions using common primers amplification. Generally, methods of the invention employ multiple steps such as template-specific hybridization, a linear amplification, partial degradation of nucleic acid, and ligation. At the end of the process the sequences of selected polynucleotides are flanked by the common sequences which can be used for exponential amplification using common primers. In some aspects the polynucleotides are associated with a barcode and the presence of the barcode is detected to measure the amount of the polynucleotide.
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10 Claims
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1. A method for amplifying an RNA target comprising:
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(a) obtaining a capture probe comprising from the 5′
end a second priming sequence, a tag sequence, a first priming sequence and a sequence complementary to the RNA target;(b) hybridizing the RNA target to the capture probe and extending the RNA using a polymerase and using the capture probe as template to form a duplex between the capture probe and the extended RNA target; (c) optionally destroying or removing the capture probe; and (d) amplifying the portion of the extended RNA comprising the first and second primer regions and the tag region. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification