METHOD FOR THE LINEAR AMPLIFICATION OF COMPLEMENTARY DNA

摘要

In one aspect, the present invention provides methods for linearly amplifying cDNA from an RNA molecule. The methods comprise the steps of: (a) synthesizing an antisense cDNA molecule using an RNA molecule as a template; (b) synthesizing an extended antisense cDNA molecule using a template-switching oligonucleotide as a template; (c) synthesizing multiple copies of sense cDNA molecules using the extended antisense cDNA strand as a template; and (d) generating double-stranded cDNA molecules using the multiple copies of sense cDNA molecules as templates. Another aspect of the invention provides methods for linearly amplifying RNA, comprising the steps o£ (a) using an RNA molecule as a template for synthesizing an antisense cDNA molecule; (b) using a template-switching oligonucleotide as a template for synthesizing an extended antisense cDNA molecule; (c) using the extended antisense cDNA molecule as a template for synthesizing multiple copies of sense cDNA molecules; (d) using the sense cDNA molecules as templates for generating double-stranded cDNA molecules; and (e) using the double-stranded cDNA molecules as templates for synthesizing RNA molecules.