m1A Sequencing

m1A sequencing is a next generation sequencing (NGS)-based method to comprehensively detect m1A, a newly discovered RNA methylation, at single nucleotide resolution. It can quickly identify the complete picture of the m1A modification in transcripts. Our experienced technical team provides you with integrated m1A sequencing service and professional data analysis.

Overview

N1-methyladenosine (m1A), occurs mainly in thousands of different gene transcripts in eukaryotic cells (from yeast to mammals). It is a post-transcriptional modification with high abundance in tRNA and rRNA in eukaryotes and most m1A methylation modifications occur in the 5'UTR region of mRNA. m1A at the first and second nucleotide of the transcript can promote mRNA translation, while m1A in the coding region can inhibit translation. Recent studies have also shown m1A plays an active and dynamic role in the initiation of translation in mammalian cells. m1A can regulate the intensity of specific translation initiation sites, thereby affecting the balance between alterative initiation sites and canonical initiation sites. m1A sequencing (m1A-seq), a technology relying on the immunoprecipitation of RNA fragments carrying m1A, provides an overview of m1A modifications. The principle is to co-incubate randomly interrupted RNA fragments with m1A RNA methylation-specific antibodies and capture methylated fragments followed by deep sequencing to provide complete maps of m1A sites in the transcriptome. m1A-seq can reveal the occurrence of m1A methylation in RNAs at high detection rate, enabling a more comprehensive understanding of transcription and translation of mRNA, structure and function of tRNA and rRNA. In particular, m1A-seq helps us to further understand the role and function of this novel epigenetic transcriptome marker.

Features

High Specificity Transcriptome-Wide Professional Team One-Step Service
Employ commercial m1A antibody that specifically captures m1A fragments. Accurate localization of m1A sites in the global transcriptome. Provide various in-depth data analysis to satisfy customer needs. Provide one-stop service from m1A enrichment, library preparation, sequencing to data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

RNA purification;
quality assessment and quantification.

Library Preparation

2. Library Preparation

RNA fragmentation;
immunoprecipitation;
m1A 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

m1A Sequencing

In-depth data analysis:

  • Statistics of m1A distribution
  • Peaks annotation
  • Evolutionary conservative analysis
  • Motif search
  • Differential binding analysis
  • Differential binding peaks annotation
  • Enrichment analysis of peaks genes
  • Correlation analysis

Sample Requirements

RNA sample: quantity ≥ 300 ug.

OD260/280 ≥ 1.8, OD260/230 ≥ 1.5, 28S:18S ≥ 1.5, RIN ≥ 7.

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. Dan D, Sigrid N, Sharon M-M, et al. The dynamic N1-methyladenosine methylome in eukaryotic messenger RNA. Nature, 2016, 25, 530 (7591): 441-446.
  2. Tserovski L, Marchand V, Hauenschild R, et al. High-throughput sequencing for 1-methyladenosine (m(1)A) mapping in RNA. Methods, 2016, 1, 107: 110-121.
  3. Safra M, Sas-Chen A, Nir R, et al. The m1A landscape on cytosolic and mitochondrial mRNA at single-base resolution. Nature, 2017, 9, 551 (7679): 251-255.
* For Research Use Only. Not for use in diagnostic procedures.


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