DNA methylation is a crucial epigenetic modification that plays a significant role in gene regulation and cellular processes. Understanding the methylation patterns in specific genomic regions is essential for unraveling the complexities of gene expression and its impact on various biological phenomena. CD Genomics, a prominent leader in the biotechnology industry, provides a cutting-edge Targeted DNA Methylation Analysis Service, including a range of methodologies, such as EpiTYPER DNA Methylation Analysis, NGS-BSP (Bisulfite Sequencing PCR), Methyl seq, CD Human Methylome Panel, and Targeted Bisulfite sequencing. With its rich expertise and state-of-the-art technologies, CD Genomics is at the forefront of offering innovative solutions for studying DNA methylation patterns with exceptional precision and accuracy.
For targeted gene/CpG site methylation detection, a flexible Targeted Bisulfite Sequencing technology has been established. CD Genomics has developed a high-throughput sequencing method called Target Bisulfite Sequencing (Target-BS), specifically designed for existing target gene panels.
Applications of Target Bisulfite Sequencing (Target-BS)
Target-BS is widely utilized for methylation biomarker screening, validation, and clinical translation at known loci.
Subsequent validation of target gene methylation.
Advantages of Target Bisulfite Sequencing Technology
Multiplex amplification of multiple genes for high-throughput detection.
Ultra-high-depth bisulfite sequencing, offering superior sensitivity and accuracy compared to traditional BSP clone sequencing, capable of detecting methylation levels below 1% in samples.
Low sample requirements, enabling multi-gene amplification with as little as 20ng genomic DNA.
Versatile applicability to genomic DNA, FFPE samples, and cell-free cfDNA.
Cost-effective with significantly lower expenses compared to existing technologies, rapid turnaround, and excellent cost-performance ratio.
Target Bisulfite Sequencing Technology Workflow
Analysis Content of Target Bisulfite Sequencing
Note: In addition to standard analysis, personalized analysis and multi-omics correlation analysis are available upon request. Please contact CD Genomics for further information.
Target Bisulfite Sequencing Sample Requirements
| Sample Type | Total Amount | Sample Concentration | Sample Integrity | Purity (OD260/280) |
|---|---|---|---|---|
| DNA | >50 ng | >10 ng/μl | No degradation and protein contamination, confirmed by gel electrophoresis | 1.8-2.0 |
| Tissue Samples | >20 mg | N/A | No degradation and protein contamination, confirmed by gel electrophoresis | 1.8-2.0 |
| Whole Blood | >0.5 ml | N/A | Should be collected using regular EDTA anticoagulant tubes, avoid using heparin anticoagulant tubes | 1.8-2.0 |
| Cell Samples | Cell count > 1x10^6 | N/A | Should be cryopreserved as live cells | 1.8-2.0 |
Recommended Data Output: 1G raw data.
Vitamin C Prevents Offspring DNA Methylation Changes Associated with Maternal Smoking in Pregnancy
Background:
Maternal smoking during pregnancy has been linked to lifelong reduction in lung function in offspring. Previous research has described DNA methylation changes associated with maternal smoking during pregnancy in various samples, such as placenta and cord blood at birth, fetal lungs, and oral epithelium and blood during childhood. This study aimed to investigate whether supplementing with vitamin C could mitigate DNA methylation changes in offspring resulting from maternal smoking during pregnancy.
Methods:
Targeted bisulfite sequencing was performed on placenta, cord blood, and oral samples. Subsequently, bisulfite amplicon sequencing was used to independently validate selected differentially methylated regions in cord blood.
Conclusion:
Among CpG sites with methylation differences of at least 10% between the placebo group and non-smokers, 69.03% showed restoration to non-smoker levels after vitamin C supplementation. Many of the restored CpG sites were associated with phenotypic outcomes enriched for low methylation CpGs. This study provides evidence that supplementing with vitamin C may reduce DNA methylation changes in offspring caused by maternal smoking during pregnancy.
The Target Bisulfite Sequencing (Target-BS) achieved an average coverage depth of 337 times for all 321 CpG sites covered in the sequencing.
Reference
- Shorey-Kendrick LE, McEvoy CT, Ferguson B, Burchard J, Park BS, Gao L, Vuylsteke BH, Milner KF, Morris CD, Spindel ER. Vitamin C Prevents Offspring DNA Methylation Changes Associated with Maternal Smoking in Pregnancy. Am J Respir Crit Care Med. 2017 Sep 15;196(6):745-755. doi: 10.1164/rccm.201610-2141OC. PMID: 28422514; PMCID: PMC5620677.