m6A (N6-methyladenosine, 6-methyl adenine) is the most common methylation modification in eukaryotic mRNA internal sequences, and can functionally regulate the eukaryotic transcriptome to affect the splicing and output of mRNA nuclear, positioning, translation and stabilization. The appearance of m6A RNA in a variety of important cell life processes means that mRNA methylation is involved in a variety of biological processes, such as stem cell differentiation, biological rhythms, etc. And it is also involved in the occurrence of many diseases, including tumors, obesity, and Infertility, etc.
IntegrateRNA provides methylated RNA immunoprecipitation sequencing (MeRIP-Seq) to provide a transcriptome-wide analysis that includes the m6A position in the RNA group. Co-immunoprecipitation of m6A-modified RNA fragments in cells with m6A-specific antibodies, and high-throughput sequencing of the enriched RNA fragments. Combined with bioinformatics analysis, m6A modifications can be systematically studied within the transcriptome.
Strategy of m6A RNA sequencing is described as bellow
Workflow of IntegrateRNA m6A RNA sequencing
- Total RNA isolation (optional, we also accept your RNA sample)
➢ Sample requirements
✧ At least 50 μg of total RNA is required. RIN value≥7.0
✧ Receiving cells, tissue samples
- Isolate RNA from total RNA and QC
- m6A immunoprecipitation with anti-m6A antibody
➢ MeRIP is based on the principle of specific binding of methylated modified bases to antibodies, based on the enrichment of methylated fragments by RNA immunoprecipitation
- IP efficiency QC via qPCR
- Sequencing library preparation
- Cluster generation
- Sequencing with Illumina next-generation sequencing platform
- Data analysis including alignment, peak recall, and annotation, motif analysis, differential methylation peak analysis, GO and path analysis.
- Using the latest next-generation sequencing technology
- Technicians are experienced and can provide experimental solutions, solve experimental problems, and analyze experimental results according to the requirements of partners.
- Unlimited range - Covers m6A methylation in any transcript region.
- High resolution - Accurately locate peaks within 100 bases of the m6A site.
- High precision - Calibrate m6A level using transcript abundance in the input controls.
- Strict quality control
- Fast turnaround time
- Comprehensive bioinformatics analysis
➢ In-depth data analysis includes
✧ Comparison, classification and functional annotation of all mapped reads
✧ Biostatistical analysis - Expression analysis, multi-parameter data analysis, length distribution, transcript copy comparison, etc.
✧ Peak calls for the entire genome
✧ Differential methylation
✧ Pattern recognition
✧ RNA differential expression analysis
✧ Correlation between RNA-Seq and m6A-Seq
✧ GO and KEGG enrichment analysis of differential genes/peak
- Expert technical support to assist in data interpretation
IntegrateRNA is a professional provider of RNA discovery, functional research, and bioinformatics analysis for assessing differential expression of RNA, identifying interacting molecules, and analyzing disease mechanisms and treatments. No matter what kind of problems encountered during your RNA research, our trained experts are ready to support you with their strong problem-solving skills and expertise. Contact us to learn more about our services. We look forward to working with you in the future.