title: Pair-barcode high-throughput sequencing for large-scale multiplexed sample analysis.
abstract: BACKGROUND
  The multiplexing becomes the major limitation of the next-generation sequencing (NGS) in application to low complexity samples. Physical space segregation allows limited multiplexing, while the existing barcode approach only permits simultaneously analysis of up to several dozen samples.
  
  
  RESULTS
  Here we introduce pair-barcode sequencing (PBS), an economic and flexible barcoding technique that permits parallel analysis of large-scale multiplexed samples. In two pilot runs using SOLiD sequencer (Applied Biosystems Inc.), 32 independent pair-barcoded miRNA libraries were simultaneously discovered by the combination of 4 unique forward barcodes and 8 unique reverse barcodes. Over 174,000,000 reads were generated and about 64% of them are assigned to both of the barcodes. After mapping all reads to pre-miRNAs in miRBase, different miRNA expression patterns are captured from the two clinical groups. The strong correlation using different barcode pairs and the high consistency of miRNA expression in two independent runs demonstrates that PBS approach is valid.
  
  
  CONCLUSIONS
  By employing PBS approach in NGS, large-scale multiplexed pooled samples could be practically analyzed in parallel so that high-throughput sequencing economically meets the requirements of samples which are low sequencing throughput demand.
authors.affiliation: State Key Laboratory of Bioelectronics, Southeast University, Nanjing, 210096, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Tu
authors.forename: Qinyu
authors.initials: Q
authors.lastname: Ge
authors.forename: Shengqin
authors.initials: S
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Beili
authors.initials: B
authors.lastname: Sun
authors.forename: Qi
authors.initials: Q
authors.lastname: Yang
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Bai
authors.forename: Zuhong
authors.initials: Z
authors.lastname: Lu
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism; D016208:Databases, Factual; D001330:Electronic Data Processing; D005260:Female; D006801:Humans; D035683:MicroRNAs / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D059467:Transcriptome
chemical_list: D035683:MicroRNAs