For the investigation of DNA methylation patterns, bisulfite sequencing PCR (BSP) is a classical method for choice. It provides detailed information on the methylation pattern of individual DNA molecules at single CpG site resolution. As a professional epigenetic research services provider, Creative BioMart offers customized BSP service of high quality, from primer design to complete analytical reports to meet your project requirements and budgets in the exploration of DNA methylation analysis.

What Is Bisulfite Sequencing PCR?

The bisulfite sequencing PCR (BSP) is a sensitive approach for directly detecting and analyzing the methylation pattern of genomic DNA, and the techniques involved include bisulfite conversion, PCR amplification and Sanger sequencing. After DNA samples are treated with bisulfite, primers are designed for PCR amplification of the target fragment, and the PCR products are cloned and sequenced. The sequenced sequence is compared with the reference sequence to determine whether the CpG site is methylated. Compared with methylation-specific PCR (MSP), this method can simultaneously detect the methylation of multiple sites, overcome the traditional MSP can only do qualitative research and able to do quantitative research, moreover, BSP only needs to design a pair of primers, and the primers at both ends of the detected sites, which reduces the risk of experimental failure due to poor primer design.

Figure 1. Schematic diagram of bisulfite sequencing PCR

Features of Bisulfite Sequencing PCR

1. High specificity, free to determine the target region to be detected, and obtain the distribution map of methylation sites in specific regions
2. High sensitivity, the accurate methylation site distribution of individual DNA molecules can be obtained by analyzing the trace genomic DNA
3. High resolution, detection of methylation status of each nucleotide

Our Advantages

1. Tolerance of multiple sample types: we currently accept DNA, cell, tissue, and formalin fixed paraffin-embedded (FFPE) tissues for our BSP service. Sample sources include human, animals, plants and microorganisms.
2. Strict quality control: we have strict quality control for each step of BSP experiment to ensure that customers receive reliable experimental data.
3. One-stop service: customers only need to provide samples, and we will complete a full BSP experiment from primer design, bisulfite conversion, PCR amplification, cloning and sequencing to data analysis. The complete report is provided to you with both raw and analyzed data, graphs, detailed protocols, and a summary of findings.

Workflow of Bisulfite Sequencing PCR at Creative BioMart

During the process of BSP service, we provide greater than 99% bisulfite conversion efficiency and professional primer design. The final report you obtained includes but not limited in PCR electropherogram, Sanger sequencing result, sequence alignment result, methylation histogram, and methylation dot plots.

Figure 2. Workflow of bisulfite sequencing PCR at Creative BioMart

Our state-of-the-art labs are staffed by some of the well-trained and experienced experts in the DNA methylation analysis field. contact us today for a free consultation with the scientific team and discover how Creative BioMart can be a valuable resource and partner for your organization.

Reference
1. Zhang Y.; et al. DNA methylation analysis by bisulfite conversion, cloning, and sequencing of individual clones. Methods Mol Biol. 2009, 507: 177-187.