Strand-specific Transcriptome Profiling with Directly Labeled RNA
News Jan 17, 2011
With lower manufacturing cost, high spot density, and flexible probe design, genomic tiling microarrays are ideal for comprehensive transcriptome studies. Typically, transcriptome profiling using microarrays involves reverse transcription, which converts RNA to cDNA. The cDNA is then labeled and hybridized to the probes on the arrays, thus the RNA signals are detected indirectly. Reverse transcription is known to generate artifactual cDNA, in particular the synthesis of second-strand cDNA, leading to false discovery of antisense RNA. To address this issue, we have developed an effective method using RNA that is directly labeled, thus by-passing the cDNA generation. This paper describes this method and its application to the mapping of transcriptome profiles.
The paper, entitled 'Strand-specific transcriptome profiling with directly labeled RNA on genomic tiling microarrays', is freely available online through BioMed Centrals Molecular Biology journal.