Pair-barcode high-throughput sequencing for large-scale multiplexed sample analysis

BMC Genomics

Methodology Article

Qinyu Ge¹  Yunfei Bai²  Shengqin Wang²  Jing Tu²  Lei Wang²  Qi Yang²  Beili Sun²  Zuhong Lu³ 

[1] Key laboratory of Child Development and Learning Science, Ministry of Education, Southeast University, 210096, Nanjing, China;State Key Laboratory of Bioelectronics, Southeast University, 210096, Nanjing, China;State Key Laboratory of Bioelectronics, Southeast University, 210096, Nanjing, China;Key laboratory of Child Development and Learning Science, Ministry of Education, Southeast University, 210096, Nanjing, China;

关键词: barcode;    next-generation sequencing;    miRNA;    breast cancer;   

DOI  :  10.1186/1471-2164-13-43

 received in 2011-04-26, accepted in 2012-01-25,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundThe 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.ResultsHere 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.ConclusionsBy 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.

【 授权许可】

Unknown   
© Tu et al; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.