Rapid and affordable genome-wide bisulfite DNA sequencing by XmaI-reduced representation bisulfite sequencing-Reference-Cited by-同舟云学术

Rapid and affordable genome-wide bisulfite DNA sequencing by XmaI-reduced representation bisulfite sequencing

Published:2017-06 Issue:6 Volume:9 Page:833-847
ISSN:1750-1911
Container-title:Epigenomics
language:en
Short-container-title:Epigenomics

Author:

Tanas Alexander S¹²,Borisova Marina E³,Kuznetsova Ekaterina B¹⁴,Rudenko Viktoria V¹,Karandasheva Kristina O¹²,Nemtsova Marina V¹⁵,Izhevskaya Vera L¹,Simonova Olga A¹,Larin Sergey S⁶⁷,Zaletaev Dmitry V¹⁴,Strelnikov Vladimir V¹²

Affiliation:

1. Epigenetics Laboratory, Research Centre for Medical Genetics, Moskvorechie Street 1, Moscow 115478, Russia

2. Molecular and Cell Genetics Chair, Pirogov Russian National Research Medical University, Moscow, Russia

3. Chromatin Biology and Proteomics Group, Institute of Molecular Biology, Mainz, Germany

4. Human Molecular Genetics Laboratory, Sechenov First Moscow State Medical University, Moscow, Russia

5. Medical Genetics Laboratory, Sechenov First Moscow State Medical University, Moscow, Russia

6. Molecular Immunology Laboratory, Dmitry Rogachev National Research Center of Pediatric Hematology, Oncology & Immunology, Moscow, Russia

7. Gene Therapy Laboratory, Institute of Gene Biology, Moscow, Russia

Abstract

Aim: To develop a reduced representation bisulfite sequencing (RRBS) approach for rapid and affordable genome-wide DNA methylation analysis. Methods: We have selected restriction endonuclease XmaI to produce RRBS library fragments. After digestion and partial fill-in DNA fragments were ligated to barcoded adapters, bisulfite converted, size-selected, and sequenced on the Ion Torrent Personal Genome Machine. XmaI-RRBS results were compared with the previously published RRBS data. Results: We have developed an XmaI-RRBS method for rapid and affordable genome-wide DNA methylation analysis, with library preparation taking only 4 days and sequencing possible within 4 h. We have also addressed several challenges in order to further improve the RRBS technology. XmaI-RRBS may be performed on degraded DNA samples and is compatible with the bench-top next-generation sequencing machines.

Publisher

Future Medicine Ltd

Subject

Cancer Research,Genetics

Link

https://www.futuremedicine.com/doi/pdf/10.2217/epi-2017-0031

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