|Premium WGBS kit MANUAL|
Whole Genome Bisulfite Sequencing (WGBS) is a single nucleotide resolution technique that a...
The Premium Whole Genome Bisulfite Sequencing (WGBS) Kit is designed to prepare single and paired-end bisulfite converted DNA libraries for sequencing using Illumina® platforms. It has also been validated for bisulfite-converted library preparation from ChIP-derived samples in order to perform ChIP-Bis-Sequencing. This kit contains specially designed enzymes and buffers needed for genome-wide bisulfite sequencing allowing for studying DNA methylation sites and their role in gene regulation. For Reduced Representation Bisulfite Sequencing (RRBS) experiments use the Diagenode’s Premium RRBS Kit.
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Figure 1: Coverage of Diagenode’s Premium WGBS Kit (A) and Premium RRBS Kit (B).
WGBS was performed using the Premium Whole Genome Bisulfite Sequencing (WGBS) kit. RRBS was performed using the Premium Reduced Representation Bisulfite Sequencing (RRBS) kit. For both, after sequencing, reads were aligned on the mm10 reference genome. Visualization of the alignment bam files on Integrative Genome Viewer (IGV) shows excellent coverage (A) of the whole genome using WGBS and (B) of the CpGs areas using RRBS.
Figure 2: ChIP-Bis-sequencing results obtained with the Diagenode’s Premium WGBS Kit
A. Chromatin Immunoprecipitation against the H3K27me3 mark was performed using the Diagenode’s iDeal ChIP-seq Kit for Histones on a Gm12878 human cell line. The library was then prepared using the MicroPlex Library Preparation Kit and sequenced. The peaks represent the distribution of the reads. B. The ChIP’d DNA was then further processed with the Premium WGBS Kit and sequenced. The distribution of the reads is shown in black. The methylated CpGs are shown in red and the unmethylated CpGs in blue. Each peak represents the DNA methylation percentage at one CpG.