rna captor a tool for rna characterization斯莱特林rna rna特征的工具.pdfVIP

RNA Captor: A Tool for RNA Characterization Christian Clepet* URGV Plant Genomics, INRA UMR1165 UEVE/CNRS ERL 8196, Evry, France Abstract Background: In the genome era, characterizing the structure and the function of RNA molecules remains a major challenge. Alternative transcripts and non-protein-coding genes are poorly recognized by the current genome-annotation algorithms and efficient tools are needed to isolate the less-abundant or stable RNAs. Results: A universal RNA-tagging method using the T4 RNA ligase 2 and special adapters is reported. Based on this system, protocols for RACE PCR and full-length cDNA library construction have been developed. The RNA tagging conditions were thoroughly optimized and compared to previous methods by using a biochemical oligonucleotide tagging assay and RACE PCRs on a range of transcripts. In addition, two large-scale full-length cDNA inventories relying on this method are presented. Conclusion: The RNA Captor is a straightforward and accessible protocol. The sensitivity of this approach was shown to be higher compared to previous methods, and applicable on messenger RNAs, non-protein-coding RNAs, transcription-start sites and microRNA-directed cleavage sites of transcripts. This strategy could also be used to study other classes of RNA and in deep sequencing experiments. Citation: Clepet C (2011) RNA Captor: A Tool for RNA Characterization. PLoS ONE 6(4): e18445. doi:10.1371/journal.pone.0018445 Editor: Juan Mata, University of Cambridge, United Kingdom Received January 3, 2011; Accepted February 28, 2011; Published April 13, 2011 Copyright: 2011 Christian Clepet. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.