Patent ID: 8329398

Claim:
A method for preparing fragmented RNA comprising a multiplicity of RNA species for gene expression analysis comprising the steps of: (a) producing randomly primed first strand cDNA from total RNA obtained from a fixed paraffin-embedded tissue (FPET) sample obtained from a human subject, using reverse transcriptase and random primers; (b) tailing said randomly primed first strand cDNA to produce tailed first strand cDNA, wherein said tailing is done using poly(A) polymerase or terminal transferase; (c) converting said tailed first strand cDNA to double stranded cDNA using a primer that anneals to said tail and contains a promoter sequence for an RNA polymerase; (d) transcribing said double stranded cDNA using said RNA polymerase to produce amplified RNA; (e) reverse transcribing said amplified RNA using a reverse transcriptase and random primers to produce a nucleic acid sample comprising amplified cDNA and amplified RNA; and (f) degrading said amplified RNA in said nucleic acid sample, thereby producing a DNA sample comprising amplified cDNA.