Amplification of RNA sequences using composite RNA-DNA primers and strand displacement

摘要

The invention provides methods for linear amplification of RNA that has an RNA sequence of interest. The methods are based on using a first DNA primer with a 3' portion that is complementary to the RNA and a 5' portion that is not complementary to the RNA in that region. The primer and an RNA-dependent DNA polymerase are used to make a DNA-RNA complex from the RNA. The RNA is cleaved from the complex with an enzyme, then a second primer and a DNA-dependent DNA polymerase are used to make double stranded DNA. The double stranded DNA is then denatured, and an RNA-DNA composite primer, a DNA-dependent DNA polymerase, and strand displacement are used to isothermally produce multiple copies of the complementary sequence to the RNA sequence of interest.