SUBTRACTIVE AMPLIFICATION

摘要

A method is provided for preferentially amplifying target RNA relative to nontarget RNA in a sample of tester RNA. According to the method, a sample of tester RNA is contacted with driver sequences which are complementary to the non-target RNA under conditions where the driver sequences hybridize to the non-target RNA. A nucleic acid primer is then extended using the target RNA as a template, forming a DNA template complementary to some or all of the target RNA. The DNA template formed using the target RNA as template is then rendered single-stranded to enable the hybridization of the DNA template to a promoter template. The DNA template is then extended using the promoter template to form an extended DNA template comprising a functional double-stranded promoter. Multiple copies of the target RNA sequence can now be transcribed from the extended DNA template through recognition of the contained functional double-stranded promoter by RNA polymerase.