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The ImProm-II Reverse Transcription System produces efficient, robust synthesis of first-strand cDNA in preparation for PCR amplification. The components of the ImProm-II Reverse Transcription System can be used to reverse transcribe RNA templates starting with total RNA, poly(A)+ mRNA or synthetic transcript RNA. The optimized reaction buffer and powerful ImProm-II Reverse Transcriptase provided in the ImProm-II System together enable robust, full-length cDNA synthesis for the reproducible analysis of rare or long messages. The cDNA synthesis conditions were formulated for standalone applications or for easy transition to gene-specific target amplification. An aliquot of the reverse transcription reaction (1-20ul) can be amplified directly using Taq DNA polymerase in coupled or uncoupled PCR.