Bisulfite library preparation is the foundational step for high-resolution DNA methylation analysis at single-base resolution. Its primary purpose is to chemically treat genomic DNA so that methylation status can be accurately decoded by next-generation sequencing (NGS). CD BioSciences provides a comprehensive, expert bisulfite library preparation service that transforms your DNA samples into sequencing-ready, high-quality libraries for precise whole-genome methylation analysis.
The basic principle of bisulfite library preparation is to chemically transform genomic DNA so that its methylation status is permanently encoded as a DNA sequence difference. This is achieved through the selective deamination of unmethylated cytosines to uracil by sodium bisulfite treatment, while methylated cytosines (5-methylcytosine) remain unchanged. Prior to this conversion, DNA is fragmented and sequencing adapters are attached. Following conversion, during PCR amplification, uracils are replicated as thymines (T), whereas methylated cytosines are replicated as cytosines (C). This creates a definitive, sequenceable map where a C-to-T transition in the final library directly indicates an originally unmethylated site, allowing for precise genome-wide methylation analysis by next-generation sequencing.
Fig.1 Principle of the bisulfite-mediated conversion of cytosine to uracil. (Grehl C, et al., 2018)
Bisulfite library preparation is a foundational and versatile technique that enables genome-wide, single-base resolution analysis of DNA methylation. By chemically converting unmethylated cytosines to uracil, it creates a sequence blueprint where methylation status is permanently encoded as a C-to-T difference. This core methodology serves as the critical upstream step for a wide array of next-generation sequencing (NGS) applications, powering diverse research and diagnostic pipelines aimed at deciphering the complex role of the epigenome in biology and disease.
Core Application Scenarios
As a leading provider in epigenetic research services, CD BioSciences offers a premium bisulfite library preparation service that combines rigorous quality control, high-efficiency conversion, and optimized protocols for challenging samples. Our expertise ensures maximum DNA integrity and minimal bias, delivering sequencing-ready libraries with exceptional reproducibility for precise, genome-wide methylation analysis. We support a wide range of downstream applications, from WGBS and RRBS to targeted panels, and provide end-to-end project support from experimental design to data-ready results.
The workflow of our service follows a stringent, multi-stage pipeline designed to preserve DNA integrity, maximize bisulfite conversion efficiency, and ensure the generation of high-quality, sequencing-ready libraries with minimal bias and maximal yield. Each step is optimized and meticulously quality-controlled to deliver reliable, publication-ready results.
Consultation & Sample QC
We begin with project alignment and rigorous assessment of your DNA's quantity, purity, and integrity to ensure it is suitable for library construction.
Library Construction
Your DNA is fragmented, end-repaired, and ligated with specialized methylated adapters to create the initial pre-conversion library.
Bisulfite Conversion
The core chemical step where unmethylated cytosines are efficiently converted to uracil, permanently encoding the methylation state into the DNA sequence.
PCR Enrichment
Converted fragments are amplified using a low-bias PCR system to generate sufficient material and incorporate indexing barcodes for multiplexing.
Final QC & Validation
Every library undergoes precise quantification and fragment size analysis to guarantee high quality and optimal performance for sequencing.
Delivery & Support
The validated, sequencing-ready libraries are delivered along with a comprehensive QC report, completing the service.
The bisulfite library preparation service from CD BioSciences serves as the cornerstone for precise, genome-wide analysis of DNA methylation (5mC). We deliver sequencing libraries of exceptional integrity and high conversion efficiency, providing a reliable foundation for your epigenetic research. If you are interested in our services, please feel free to contact us for more details and quotation information of related services.
Reference 1. Grehl C, Kuhlmann M, Becker C, et al. How to design a whole-genome bisulfite sequencing experiment[J]. Epigenomes, 2018, 2(4): 21.
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