--- title: "Bisulfite Sequencing" search_description: "IGATech’s bisulfite sequencing (BS-Seq) services enable precise DNA methylation analysis, ideal for epigenetics research in gene regulation, disease mechanisms, and environmental studies. Achieve accurate methylation profiling with our advanced solutions." published: 2024-11-10 22:00:29.258825+00:00 locale: "it" --- # Bisulfite Sequencing [Request a quote](https://igatechnology.com/igatech/contacts/#contact-form) ## Technologies ![Nanopore Sequencing](/media/images/1.width-500.jpg) ### Nanopore Sequencing Nanopore sequencing is empowering many applications by the ability to analyze DNA, RNA and their modifications with no limitation on sequence length. This is of particular importance when the goal is to resolve hard-to-sequence or repetitive regions that have so far hampered the possibility to decode structural and epigenetic variation. Also, direct RNA sequencing and full-length cDNA sequencing are providing unprecedented insight into the real strucure and abundance of transcripts isoforms. ![NovaSeq X Plus](/media/images/Illumina_NovaSeq_XPlus_Left_561x375.width-500.png) ### NovaSeq X Plus The NovaSeq X Plus is a powerful and scalable ultra-high-throughput sequencing system that supports the broadest range of applications and study sizes. Unrivaled data quality using Illumina's proven SBS chemistry has made the NovaSeq X Plus the instrument of choice for all major genome centers and leading institutions throughout the world. Producing up to 25 billion reads \(7.5 Tbp\) per flowcell, yielding up to 120 human genomes in about 2 days at full capacity. ![Bisulfite treatment](/media/images/Bisulfite_treatment.width-500.png) ### Bisulfite treatment Bisulfite treatment allows to explore methylation levels of genomic DNA. Non-methylated cytosine are converted to uracil and then to thymine. Sequencing and mapping to adequate read depth allows to detect the level of 5mC \(5-methylcytosine\) on DNA molecules either on CpG, CHG or CHH contexts for both strands of the DNA molecules. See [BS-Seq - RRBS-Seq](https://igatechnology.com/genomics-research-services/bs-seq/). ![AVITI Element Biosciences](/media/images/0.width-500.png) ### AVITI Element Biosciences The AVITI System by Element Biosciences is a benchtop sequencer designed for high performance, cost efficiency, and flexibility. It features dual independent flow cells for parallel or independent operation, enabling flexibility in throughput from low to full. The system utilizes Avidity Sequencing™ technology, ensuring over 90% of reads exceed Q30 quality, with minimal errors, low duplication rates, and reduced AT/GC bias. It supports easy data conversion to FASTQ files, offers compatibility with numerous assays, and provides cost-effective sequencing. This is our ideal companion for dedicated and fast-turnaround run set-up. For more detailed information, you can visit their website directly at [https://www.elementbiosciences.com/products/aviti](https://www.elementbiosciences.com/products/aviti). ![agouti](/media/images/agouti.width-1500.png) ## SOLVING THE GENOME'S SECOND CODE DNA methylation is a fundamental mechanism of the epigenetic regulation of gene activity. It has been shown that DNA methylation plays an important role in a wide variety of biological processes, including silencing of transposable elements, stem cell differentiation, embryonic development, genomic imprinting and inflammation. In addition, alteration of methylation patterns has been identified in many diseases, inflammation and neurological disorders. By combining bisulfite treatment of genomic DNA with Next-Gen sequencing it is possible to sensitively measure cytosine methylation on a genome-wide scale within specific sequence contexts. However, the analysis of each methylated cytosin in the genome might be quite expensive. By using restriction enzymes and bisulfite sequencing it is possible to enrich for the areas of the genome that have higher CpG content. This approach, termed RRBS-seq \(Reduced Representation Bisulfite Sequencing\), reduces the amount of nucleotides needed to be sequenced to 1% of genome size, allowing for a cost-effective single-base-pair resolution of methylated cytosines. The third alternative is targeted bisulfite sequencing, which is able to specifically capture selected genomic regions of interest associated with a disease or phenotype. IGATech is adopting conversion protocol to analyze both 5mC and 5hmC in the same experiment. ![5mc-5hmC](/media/images/5mc-5hmC.width-1500.jpg) . ### WHOLE GENOME BS-seq \(WGBS-seq\) The process of bisulfite treatment denatures 50 ng of genomic DNA into single-stranded DNA \(ssDNA\). The protocol converts bisulfite-treated, ssDNA into an Illumina sequencing library. Post-bisulfite conversion method ensures that all ssDNA fragments are captured during the procedure, eliminating sample loss. ### REDUCED REPRESENTATION BS-seq \(RRBS-seq\) RRBS is used to generate single base resolution DNA methylation \(5-methylC\) information across a genomic sample, starting form as little as 100 ng of high-quality genomic DNA. The current approach utilizes the methylation insensitive restriction enzymes *MspI* for mammals and *TaqI* for plant species. The introduction of integrated molecular tag during the sample processing enables the removal of non-unique reads from the dataset. ### TARGETED BISULFITE SEQUENCING \(human only\) Bisulfite conversion sequencing can be done with targeted methods such as target enrichment. The protocols used are fully customized and enable the targeting of selected genomic regions from bisulfite treated genomic DNA in a single workflow, in order to identify specific regions in the genome for methylation variation assessment. As final analysis, it is possible to target selected human genomic regions from bisulfite treated genomic DNA in a single workflow. Our team is always available to consult with you on study design to ensure correct sequencing and bioinformatics strategies are used to meet your goals. ## BIOINFORMATICS SERVICES **We offer state-of-art analysis pipelines for WGBS-seq, RRBS-seq and Targeted BS-seq. The workflow includes:** - Quality control - Post-sequencing estimation of conversion rate using Lambda spike-in - Alignment and deduplication - Methylation calling at single cytosine level - Whole genome methylation statistics \(including distribution of methylation in the CG, CHG and CHH context in plants\) - Visualization of methylation distribution across the genome using CIRCOS image - Identification of Differentially Methylated Regions \(DMRs\) between samples - Annotation of DMRs ## Terms and Conditions - [Privacy Information.pdf](/documents/12/Privacy_Information_rev00.pdf) - [Human samples clearance.pdf](/documents/13/__Human-samples_clearance.pdf) - [Shipping and Packaging Guidelines.pdf](/documents/9/SHIPPING_AND_PACKAGING_GUIDELINES.pdf) - [rev05 Terms and Conditions.pdf](/documents/137/__M12_01_rev05_Terms_and_Conditions.pdf)