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SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states

Authors :
Jinke Wang
Wei Dai
Jian Wu
Ling Wu
Source :
BMC Genomics, Vol 19, Iss 1, Pp 1-12 (2018), BMC Genomics
Publication Year :
2018
Publisher :
BMC, 2018.

Abstract

Background Next-generation sequencing (NGS) is fundamental to the current biological and biomedical research. Construction of sequencing library is a key step of NGS. Therefore, various library construction methods have been explored. However, the current methods are still limited by some shortcomings. Results This study developed a new NGS library construction method, Single strand Adaptor Library Preparation (SALP), by using a novel single strand adaptor (SSA). SSA is a double-stranded oligonucleotide with a 3′ overhang of 3 random nucleotides, which can be efficiently ligated to the 3′ end of single strand DNA by T4 DNA ligase. SALP can be started with any denatured DNA fragments such as those sheared by Tn5 tagmentation, enzyme digestion and sonication. When started with Tn5-tagmented chromatin, SALP can overcome a key limitation of ATAC-seq and become a high-throughput NGS library construction method, SALP-seq, which can be used to comparatively characterize the chromatin openness state of multiple cells unbiasly. In this way, this study successfully characterized the comparative chromatin openness states of four different cell lines, including GM12878, HepG2, HeLa and 293T, with SALP-seq. Similarly, this study also successfully characterized the chromatin openness states of HepG2 cells with SALP-seq by using 105 to 500 cells. Conclusions This study developed a new NGS library construction method, SALP, by using a novel kind of single strand adaptor (SSA), which should has wide applications in the future due to its unique performance. Electronic supplementary material The online version of this article (10.1186/s12864-018-4530-3) contains supplementary material, which is available to authorized users.

Details

Language :
English
ISSN :
14712164
Volume :
19
Issue :
1
Database :
OpenAIRE
Journal :
BMC Genomics
Accession number :
edsair.doi.dedup.....2647f8a67cf0a9d0f4e32f2bb5a2d0ce
Full Text :
https://doi.org/10.1186/s12864-018-4530-3