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SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states
- 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.
- Subjects :
- 0301 basic medicine
Library construction
Library
lcsh:QH426-470
Computer science
Library preparation
lcsh:Biotechnology
Genomics
Computational biology
HeLa
Preparation method
chemistry.chemical_compound
03 medical and health sciences
lcsh:TP248.13-248.65
Tn5
Genetics
Nucleotide
Denaturation (biochemistry)
Genomic library
Throughput (business)
Salp
chemistry.chemical_classification
biology
Oligonucleotide
Methodology Article
biology.organism_classification
Chromatin
Chromatin state
lcsh:Genetics
030104 developmental biology
chemistry
Single strand adaptor
SALP-seq
DNA microarray
DNA
Biotechnology
Subjects
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