scalable transcriptome preparation for massive parallel sequencing可伸缩的转录组准备大规模并行测序.pdfVIP

Scalable Transcriptome Preparation for Massive Parallel Sequencing 1. 1. 2 1 Henrik Stranneheim , Beata Werne , Ellen Sherwood , Joakim Lundeberg * 1 Science for Life Laboratory, Division of Gene Technology, School of Biotechnology, Royal Institute of Technology (KTH), Solna, Sweden, 2 Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Solna, Sweden Abstract Background: The tremendous output of massive parallel sequencing technologies requires automated robust and scalable sample preparation methods to fully exploit the new sequence capacity. Methodology: In this study, a method for automated library preparation of RNA prior to massively parallel sequencing is presented. The automated protocol uses precipitation onto carboxylic acid paramagnetic beads for purification and size selection of both RNA and DNA. The automated sample preparation was compared to the standard manual sample preparation. Conclusion/Significance: The automated procedure was used to generate libraries for gene expression profiling on the Illumina HiSeq 2000 platform with the capacity of 12 samples per preparation with a significantly improved throughput compared to the standard manual preparation. The data analysis shows consistent gene expression profiles in terms of sensitivity and quantification of gene expression between the two library preparation methods. Citation: Stranneheim H, Werne B, Sherwood E, Lundeberg J (2011) Scalable Transcriptome Preparation for Massive Parallel Sequencing. PLoS ONE 6(7): e21910. doi:10.1371/journal.pone.0021910 Editor: Vladimir Brusic, Dana-Farber Cancer Institute, United States of America Received December 17, 2010; Accepted June 14, 2011; Publ