Evaluation of the whole genome amplification of bisulfite modified DNA
ABSTRACT: We compared genom-wide DNA methylation profiles between whole genome amplified bisufite-modified DNA and non-amplified DNA. 3 samples were bisulfite-converted, non-amplified DNA (technical triplicate). 6 samples were amplified with EpiTect® Whole Bisulfitome from 10ng or 50ng of bisulfite-converted DNA. All samples were hybridized to the Illumina Infinium HumanMethylation450.
Project description:By depleting CGGBP1 in normal human fibroblasts and by performing genome-wide sequencing (with and without bisulfite conversion) we show that upon CGGBP1 depletion cytosine methylation increases significantly at repeat regions. Using Pacbio sequencing of Alu and LINE-1 repeats amplified genome-wide from bisulfite converted DNA, we further establish the cytosine methylation-inhibitory functions of CGGBP1. Overall design: Control shmiR and CGGBP1 shmiR genomic DNA (both with or without sodium bisulfite conversion) used for Illumina sequencing experiments. Further bisulfite converted samples were used as templates to amplify Alu-SINEs and L1-LINEs genome wide and used for PacBio sequencing experiment. We have sequenced the human genomic DNA (no experimental enrichment, so no modification of BAM files during analyses) and used the BAM files, the most analyzed files, to calculate the genome composition statistics, such as base composition bias etc. The differences in base composition (C->T drift upon CGGBP1-depletion) changes are our key findings.
Project description:Genome wide DNA methylation in blood derived DNA from individuals with deletion or duplication of 7q11.23 Bisulfite converted DNA was hybridized onto the Illumina Infinium HumanMethylation450 BeadChip array
Project description:Whole genome bisulfite sequencing of MDCK cells, before and after TGFB-induced epithelial-mesenchymal transition Sequencing of bisulfite converted DNA from MDCK cells untreated, and after a 30 days treatment with TGF beta
Project description:The ability to predict the type of tissues or cells from molecular profiles of crime scene samples has practical implications in forensics. In order to identify body fluid-specific DNA methylation changes, genome-wide DNA methylation profiling was carried out for body fluids including menstrual blood and vaginal fluid obtained from 3 female donors aged 19, 27 and 38 years. The Illumina Infinium 450k Human DNA methylation Beadchip was used to obtain DNA methylation profiles across approximately 450,000 CpGs in bisulfite converted DNA. Samples included 3 of each vaginal fluid and menstrual bloods collected on the first, second and third days of menstrual bleeding. Bisulfite converted DNA from the 12 samples were hybridised to the Illumina Infinium 450k Human Methylation Beadchip
Project description:Bisulfite conversion and whole genome-single base next generation sequencing of DNA from a single iPSC clone (CMC28). This method provides exceptional depth of the sequenced methylome. Bisulfite converted DNA from a single iPSC clone (CMC28), and get its high-throughput sequence data with Illumina.
Project description:Genome-scale DNA methylation was analyzed in a cohort of paragangliomas to identify DNA methylation changes. Bisulfite converted DNA from 22 fresh frozen paragangliomas and 2 normal adrenal medulla samples were hybridized to Illumina HumanMethylation450 BeadChips.
Project description:Genome-scale DNA methylation was analyzed in a cohort of ACC and ACA to identify DNA methylation changes. Bisulfite converted DNA from 51 fresh frozen ACC and 30 ACA samples were hybridized to Illumina HumanMethylation27 BeadChips.
Project description:We applied the solution hybrid selection approach to the enrichment of CpG islands (CGIs) and promoter sequences from the human genome for targeted high-throughput bisulfite sequencing. A single lane of Illumina sequences allowed accurate and quantitative analysis of 1 million CpGs in more than 21,408 CGIs and 15,946 transcriptional regulatory regions. More than 85% of capture probes successfully yielded quantitative DNA methylation information of targeted regions. In this study, we generated genome-wide, single-base resolution DNA methylation maps in three of the most commonly used breast cancer cell lines.Differentially methylated regions (DMRs) were identified in the 5?-end regulatory regions, as well as the intra- and intergenic regions, particularly in the X chromosome among the three cell lines. The single CpG resolution methylation maps of many known tumor suppressor genes were also established in the three cell lines. Here we present a novel approach that combines solution-phase hybrid selection and massively parallel bisulfite sequencing to profile DNA methylation in targeted CGI and promoter regions. We designed 51,466 single strand DNA oligonucleotides (160-mer) which target 23,441 CGIs and the transcription start sites of 19,369 known genes in the human genome. The synthetic long DNA oligonucleotides were converted into biotinylated RNA probes for solution-phase hybridization capture of target DNA. The captured genomic DNA was treated with sodium bisulfite, amplified by PCR and sequenced using Illumina GA IIx sequencer.
Project description:Genomewide DNA methylation profiling of monocyte isolated from humna blood, immature and mature dendritic cells generated ex vivo. The Illumina Infinium HumanMethylation450 beadchips were used to generate DNA methylation profiles. Bisulfite converted DNA were hybridized to the Illumina Infinium HumanMethylation450 beadchips