Method of amplifying DNA from RNA in a sample

摘要

A method of amplifying RNA in a sample and a method of amplifying a pool of RNA is provided. Gene loss and amplification bias may be reduced using the methods. Also, directionality may be preserved in the amplified RNA. The methods may be applied in sequence analysis as well as in general molecular diagnostic areas.

主权项

1. A method of amplifying RNA in a sample, the method comprising:
ligating a single-stranded adapter nucleic acid to any one of two ends of RNA in a sample to thereby form an adapter nucleic acid-RNA fusion construct, wherein the adapter nucleic acid-RNA construct consists of the RNA and the single-stranded adapter nucleic acid bound to one end of the RNA; self-ligating the adapter nucleic acid-RNA construct to thereby form a circular adapter nucleic acid-RNA fusion construct; and amplifying the RNA by incubating the circular adapter nucleic acid-RNA fusion construct together with a first primer and a second primer to thereby form an amplification product, wherein when the single-stranded adapter nucleic acid is ligated to the RNA, the first primer comprises a nucleotide sequence that is the same as a continuous nucleotide sequence in the adapter nucleic acid and the second primer comprises a nucleotide sequence that is complementary to a continuous nucleotide sequence in the adapter nucleic acid.