COMPOSITIONS AND METHODS FOR THE ANALYSIS OF DEGRADED NUCLEIC ACIDS

摘要

The invention relates to compositions and methods for gene expression analysis. In some embodiments, the invention provides compositions and methods for amplifying targets in a degraded nucleic acid sample. In some embodiments, the invention provides methods for determining the quality of nucleic acids (e.g., the degree of degradation) in a nucleic acid sample. The invention also provides methods for producing a gene expression profile from a degraded RNA sample.

主权项

1. A method for amplifying members of a population of degraded nucleic acids in a sample, the method comprising:
a) providing a sample comprising said population of degraded nucleic acids; b) providing target primer pairs, wherein
i) each target primer pair comprises a forward target primer and a reverse target primer;ii) said forward and reverse target primers each comprise (A) a target-specific nucleotide sequence that is complementary to a nucleotide subsequence of at least one member of the population of degraded nucleic acids, and (B) at least one universal priming sequence, wherein the universal priming sequence is 5′ relative to the target-specific sequence; and c) annealing said target primer pairs to their cognate degraded nucleic acid; d) enzymatically producing a plurality of products corresponding to subsequences of said cognate degraded nucleic acids; e) enzymatically amplifying said plurality of products using at least one universal primer to produce a plurality of target amplicons, wherein said universal primer comprises nucleotide sequence that is complementary to said universal priming sequence, thereby amplifying members of a population of degraded nucleic acids.