title: High concordance between Illumina HiSeq2500 and NextSeq500 for reduced representation bisulfite sequencing (RRBS).
abstract: Reduced representation bisulfite sequencing (RRBS) provides an efficient method for measuring DNA methylation at single base resolution in regions of high CpG density. This technique has been extensively tested on the HiSeq2500, which uses a 4-colour detection method, however it is unclear if the method will also work on the NextSeq500 platform, which employs a 2-colour detection system. We created an RRBS library and sequenced it on both the HiSeq2500 and NextSeq500, and found no significant difference in the base composition of reads derived from either machine. Moreover, the methylation calls made from the data of each instrument were highly concordant, with methylation patterns across the genome appearing as expected. Therefore, RRBS can be sequenced on the Nextseq500 with comparable quality to that of the HiSeq2500. All sequencing data are deposited in the GEO database under accession number GSE87097.
authors.affiliation: Molecular Medicine Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.
authors.forename: Danqing
authors.initials: D
authors.lastname: Yin
authors.affiliation: Molecular Medicine Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia; Department of Medical Biology, Parkville, Victoria 3010, Australia; School of Mathematics and Statistics, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Ritchie
authors.affiliation: The Australian Genome Research Facility, Melbourne, Parkville, Victoria 3052, Australia.
authors.forename: Jafar S
authors.initials: JS
authors.lastname: Jabbari
authors.affiliation: Molecular Medicine Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.
authors.forename: Tamara
authors.initials: T
authors.lastname: Beck
authors.affiliation: Molecular Medicine Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia; Department of Medical Biology, Parkville, Victoria 3010, Australia.
authors.forename: Marnie E
authors.initials: ME
authors.lastname: Blewitt
authors.affiliation: Molecular Medicine Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia; Department of Medical Biology, Parkville, Victoria 3010, Australia.
authors.forename: Andrew
authors.initials: A
authors.lastname: Keniry
keywords: Bisulfite sequencing; HiSeq2500; NextSeq500; RRBS