Adapters for ligation to RNA in an RNA library with reduced bias

摘要

An improved adapter design for ligation to target RNA in a library is described which enables target RNAs that would normally be under-represented or not represented at all in a cDNA library or a PCR product of the cDNA library, to be represented with average frequency.

主权项

1. A set of reagents for producing adaptor-ligated RNA, comprising:
(a) a population of 3′ polynucleotide adaptors each having a 3′ end and a 5′ end, wherein each adaptor comprises:
(i) a first non-random sequence of at least seven defined nucleotides at the 5′ end,(ii) a first random nucleotide sequence located between the 3′ end and the first defined nucleotide sequence,(iii) a third non-random sequence of at least 5 defined nucleotides, and(iv) a modified nucleotide at the 3′ end,and wherein the first random nucleotide sequence varies in the population of 3′ polynucleotide adaptors; (b) a 5′ polynucleotide adapter, the 5′ adapter having a 3′ end and a 5′ end and comprising a second non-random sequence of at least seven contiguous nucleotides that is complementary to the first non-random sequence of (a)(i); and (c) a ligase capable of ligating RNA.