On-flowcell reverse transcription facilitates amplification-free, strand-specific, transcriptome sequencing

We report an alternative approach to transcriptome sequencing for the Illumina Genome Analyzer, in which the reverse transcription reaction takes place on the flowcell. No amplification is used during the library preparation, so PCR biases and duplicates are avoided, and because the template is mRNA rather than cDNA, the resulting sequences are necessarily strand-specific. The method is compatible with paired- or single-ended sequencing.