title: Scalable transcriptome preparation for massive parallel sequencing.
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.
  
  
  METHODS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Science for Life Laboratory, Division of Gene Technology, School of Biotechnology, Royal Institute of Technology (KTH), Solna, Sweden.
authors.forename: Henrik
authors.initials: H
authors.lastname: Stranneheim
authors.forename: Beata
authors.initials: B
authors.lastname: Werne
authors.forename: Ellen
authors.initials: E
authors.lastname: Sherwood
authors.forename: Joakim
authors.initials: J
authors.lastname: Lundeberg
mesh_terms: D001331:Automation; D045744:Cell Line, Tumor; D011232:Chemical Precipitation; D018076:DNA, Complementary / Q000096:biosynthesis; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary; D012334:RNA, Neoplasm