Compositions for amplification of RNA sequences using composite primers

摘要

The invention provides methods for isothermal amplification of RNA. The methods are particularly suitable for amplifying a plurality of RNA species in a sample. The methods employ a composite primer, a second primer and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In another aspect, the methods employ a single primer (which is a composite primer) and strand displacement to generate multiple copies of DNA products comprising sequences complementary to an RNA sequence of interest. In some embodiments, a transcription step is included to generate multiple copies of sense RNA of an RNA sequence of interest. The methods are useful for preparation of nucleic acid libraries and substrates for analysis of gene expression of cells in biological samples. The invention also provides compositions and kits for practicing the amplification methods, as well as methods which use the amplification products.

主权项

1. A composition comprising at least one RNA template and a plurality of primers, wherein the primers are composite primers that comprise a 5′ RNA portion and a 3′ DNA portion, wherein the 3′ DNA portion comprises a random sequence, and wherein the random sequence in at least one of the plurality of primers is complementary to a sequence in the at least one RNA template.