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 PMID:29544553  

Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data.

Nelly Olova | Felix Krueger | Simon Andrews | David Oxley | Rebecca V Berrens | Miguel R Branco | Wolf Reik
Genome biology | 2018

Whole-genome bisulfite sequencing (WGBS) is becoming an increasingly accessible technique, used widely for both fundamental and disease-oriented research. Library preparation methods benefit from a variety of available kits, polymerases and bisulfite conversion protocols. Although some steps in the procedure, such as PCR amplification, are known to introduce biases, a systematic evaluation of biases in WGBS strategies is missing.

Pubmed ID: 29544553

Research resources used in this publication

None found

Antibodies used in this publication

None found

Associated grants

  • Agency: Biotechnology and Biological Sciences Research Council, United Kingdom
    Id: BB/K010867/1
  • Agency: Sir Henry Dale Fellowship, International
    Id: 101225/Z/13/Z)
  • Agency: Wellcome Trust, United Kingdom
    Id: 095645/Z/11/Z
  • Agency: EU BLUEPRINT, International
    Id: 282510
  • Agency: Wellcome Trust, United Kingdom
  • Agency: EU EpiGeneSys, International
    Id: 257082

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ATCC (tool)

RRID:SCR_001672

Global nonprofit biological resource center (BRC) and research organization that provides biological products, technical services and educational programs to private industry, government and academic organizations. Its mission is to acquire, authenticate, preserve, develop and distribute biological materials, information, technology, intellectual property and standards for the advancement and application of scientific knowledge. The primary purpose of ATCC is to use its resources and experience as a BRC to become the world leader in standard biological reference materials management, intellectual property resource management and translational research as applied to biomaterial development, standardization and certification. ATCC characterizes cell lines, bacteria, viruses, fungi and protozoa, as well as develops and evaluates assays and techniques for validating research resources and preserving and distributing biological materials to the public and private sector research communities.

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SeqMonk (tool)

RRID:SCR_001913

Software tool to visualize and analyse high throughput mapped sequence data.

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Ensembl (tool)

RRID:SCR_002344

Collection of genome databases for vertebrates and other eukaryotic species with DNA and protein sequence search capabilities. Used to automatically annotate genome, integrate this annotation with other available biological data and make data publicly available via web. Ensembl tools include BLAST, BLAT, BioMart and the Variant Effect Predictor (VEP) for all supported species.

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Bismark (tool)

RRID:SCR_005604

Software tool to map bisulfite converted sequence reads and determine cytosine methylation states. Flexible aligner and methylation caller for Bisulfite-Seq applications. Used to map bisulfite treated sequencing reads to genome of interest and perform methylation calls in single step.

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QUMA (tool)

RRID:SCR_010907

You can easily align, visualize and quantify bisulfite sequence data for CpG methylation analysis.

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New England Biolabs (tool)

RRID:SCR_013517

An Antibody supplier

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FastQC (tool)

RRID:SCR_014583

Quality control software that perform checks on raw sequence data coming from high throughput sequencing pipelines. This software also provides a modular set of analyses which can give a quick impression of the quality of the data prior to further analysis.

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