m6A Profiling

N6-methyladenosine (m6A) profiling is a next generation sequencing (NGS)-based method to comprehensively detect m6A, a common and abundant RNA methylation modification that exists in various transcripts. Our experienced technical team provides you with multiple m6A detection services, including MeRIP-seq, PA-m6A and miCLIP-m6A. Meantime, we provide professional in-depth data analysis to meet customer needs.

Overview

Interest in RNA modification has grown rapidly in recent years, including m6A. m6A is the most common post-transcriptional modification of eukaryotic mRNA, accounting for 80% of RNA methylation modification, and is mainly present in 3’UTR. m6A is a dynamic and reversible modification process, which is regulated by methyl-transferase complex, demethylase and corresponding receptors. Existing studies have shown that m6A plays an important role in accelerating mRNA metabolism and translation in cells, as well as in cell differentiation, embryonic development, and stress response, is a hot spot of epigenetic transcriptome research. With the development of high-throughput sequencing technology, the resolution and sensitivity of methylation detection have been greatly improved. Our company provides multiple m6A detection methods, including MeRIP-seq, miCLIP-m6A and PA-m6A. MeRIP-seq is a method that combines immunoprecipitation with NGS technology to quickly and efficiently detect genomic m6A methylation, but it cannot achieve single-base resolution. miCLIP-m6A is a method that cross-links mRNA with m6A antibody and uses single-nucleotide resolution to accurately locate m6A in transcriptome. And PA-m6A is a method that uses RNase T1 to digest RNA to approximately 30nt for efficient sequencing. This method also has the characteristic of single-base resolution. We provide multiple analysis strategies for customers to choose, which can comprehensively analyze the distribution and changes of m6A in transcriptome, help researchers deeply understand the role of RNA modification in life and disease, and promote accurate diagnosis and treatment of diseases.

Features

Application High Efficiency Bioinformatics Analysis Flexibility
This method can profile and quantify m6A in small-RNA species and m6Am. Adopts carefully optimized experimental procedure, achieving high efficiency and specificity. Our integrated bioinformatics pipeline can be tailored to suit your project. One-stop solution from sample QC, library construction, to sequencing and data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

RNA purification;
quality assessment and quantification.

Library Preparation

2. Library Preparation

Poly(A) tail RNA selection;
m6A library preparation.

Sequencing

3. Sequencing

Illumina HiSeq;
PE 50/75/100/150.

Data Analysis

4. Data Analysis

Visualize and preprocess results, and perform custom bioinformatics analysis.

Bioinformatics Analysis Pipeline

m6A Profiling

In-depth data analysis:

  • Statistics of m6A distribution
  • Peak calling on whole genome
  • Peaks annotation
  • Transcriptome-wide profiling of m6A methylation
  • Differential binding analysis
  • Motif identification
  • GO&KEGG enrichment analysis of different peaks
  • Identify m6A and m6Am in small RNA
  • Clustering analysis and enrichment analysis

Sample Requirements

Please make sure that the RNA is not significantly degraded.

Sample storage: RNA can be dissolved in ethanol or RNA-free ultra-pure water and stored at -80°C. RNA should avoid repeated freezing and thawing.

Shipping Method: When shipping RNA samples, the RNA sample is stored in a 1.5 mL Eppendorf tube, sealed with sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.

Deliverable: FastQ, BAM, coverage summary, QC report, custom bioinformatics analysis.

References:

  1. Mauer J, Luo X, Blanjoie A, et al. Reversible methylation of m6Am in the 5 cap controls mRNA stability. Nature, 2017, 541: 371-375.
  2. Linder B, Grozhik A V, Olarerin-George A O, et al. Single-nucleotide-resolution mapping of m6A and m6Am throughout the transcriptome. Nat Methods, 2015, 12: 767-772.
  3. Dominissini D, Moshitch-Moshkovitz S, Schwartz S, et al. Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature, 2012, 485(7397): 201-206.
* For Research Use Only. Not for use in diagnostic procedures.


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