Methods for linearly amplifying mRNA to produce antisense RNA are provided.
In the subject methods, mRNA is converted to double-stranded cDNA using a promoter-primer
having a poly-dT primer site linked to a promoter sequence so that the resulting
double-stranded cDNA is recognized by an RNA polymerase. The resultant double-stranded
cDNA is then transcribed into antisense RNA in the presence of a reverse transcriptase
that is rendered incapable of RNA-dependent DNA polymerase activity during this
transcription step. The subject methods find use a variety of different applications
in which the preparation of linearly amplified amounts of antisense RNA is desired.
Also provided are kits for practicing the subject methods.
