GLOBAL AMPLIFICATION USING RANDOM PRIMING BY A COMPOSITE PRIMER
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Abstract
The invention relates to the field of polynucleotide amplification. More particularly, the invention provides methods, compositions and kits for amplification of (i.e., making multiple copies of) a multiplicity of different polynucleotide template sequences using a randomly primed RNA/DNA composite primer.
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Citations
227 Claims
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1-190. -190. (canceled)
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191. A kit comprising:
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(a) a first composite primer comprising a 5′
segment and a 3′
segment, wherein the 5′
segment comprises RNA and the 3′
segment comprises DNA, wherein said composite primer is designed to randomly hybridize to a multiplicity of sites on a template polynucleotide; and(b) an auxiliary primer comprising DNA, wherein said auxiliary primer is designed to randomly hybridize to a multiplicity of sites on said template polynucleotide. - View Dependent Claims (193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 209, 210, 211, 212, 213, 214, 215, 220, 221, 222)
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192. A kit comprising:
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(a) a first composite primer comprising a 5′
segment and a 3′
segment, wherein the 5′
segment comprises RNA and the 3′
segment comprises DNA, wherein said composite primer is designed to randomly hybridize to a multiplicity of sites on a template polynucleotide comprising DNA; and(b) an amplification primer, wherein said amplification primer is a composite primer comprising an RNA portion and a 3′
DNA portion, wherein the amplification primer comprises some of the sequence of the first primer, and wherein the first primer and the amplification primer are different primers. - View Dependent Claims (208, 216, 217, 218, 219)
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223. A kit comprising:
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(a) a first composite primer comprising a 5′
segment and a 3′
segment, wherein the 5′
segment comprises RNA and the 3′
segment comprises DNA, wherein said composite primer is a population of primers that are designed to hybridize to a multiplicity of sites on a template polynucleotide; and(b) an auxiliary primer comprising DNA, wherein said auxiliary primer is designed to hybridize to a multiplicity of sites on said template polynucleotide. - View Dependent Claims (225, 226, 227)
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224. A kit comprising:
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(a) a first composite primer comprising a 5′
segment and a 3′
segment, wherein the 5′
segment comprises RNA and the 3′
segment comprises DNA, wherein said composite primer is a population of primers that are designed to hybridize to a multiplicity of sites on a template polynucleotide comprising DNA; and(b) an amplification primer, wherein said amplification primer is a composite primer comprising an RNA portion and a 3′
DNA portion, wherein the amplification primer comprises some of the sequence of the first primer, and wherein the first primer and the amplification primer are different primers.
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Specification