O8G-Seq

O8G RNA oxidative modification, also known as 8-oxoguanine oxidation, is a new type of RNA modification. O8G-Seq is another hot spot technology in epitranscriptomics after m6A-Seq. O8G is a kind of RNA modification produced by reactive oxygen species attacking guanine in mRNA in cells, which can pair with adenine and induce guanine-thymine (G>T) mutation. This process is a modification in pathophysiology that leads to disease phenotypes through reactive oxygen species oxidation.

Overview

CD Genomics offers O8G RNA oxidation detection by O8G-IP-Seq technology. The technical principle is as follows: Co-incubate O8G-specific antibodies with randomly interrupted RNA fragments, grabbing the O8G-modified fragments for sequencing; A control (Input) sample is sequenced in parallel, and the control sample only contains interrupted RNA fragments and is incubated without O8G-specific antibody. The control sample is used to eliminate the background of non-specifically captured O8G fragments; Compare the sequence fragments of the co-immunoprecipitation (IP) sample with the control sample (Input), locate the O8G oxidation modification site on the transcriptome, and calculate the degree of O8G oxidation in the sample and its RNA expression level according to the RNA-seq data.

CD Genomics not only performs O8G whole-transcriptome sequencing, which can simultaneously detect O8G oxidized circular RNA, lncRNA, mRNA and more, but also detects the O8G oxidation of RNA in mRNA, lnRNA and cricRNA, respectively. With professional bioinformatics analysis platform, we can further provide high-precision O8G oxidation mapping to help customers study the biological functions and potential mechanisms of O8G oxidation. Featured analyses include: identification of O8G RNA oxidation-enriched regions, O8G RNA oxidation region annotation, GO analysis and KEGG signaling pathway analysis of O8G RNA oxidation.

Features

Any Species Full Transcriptome Coverage High Specificity Professional Bioinformatics
This method can detect RNA O8G oxidation of any species, from animal to plant. Whole-transcriptome wide RNA O8G oxidation profiling. Specific enrichment and detection of O8G oxidized RNA fragments. Strong bioinformatic team, professional O8G oxidation data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

RNA purification; quality assessment and quantification.

Library Preparation

2. Library Preparation

RNA fragmentation; crosslinking reaction; PCR amplification.

Sequencing

3. Sequencing Platform

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

Data Analysis

4. Data Analysis

Preprocess and visualize results, and perform custom bioinformatics analysis.

O8G-Seq

Bioinformatics Analysis Pipeline

In-depth data analysis:

  • Quality control of sequencing results
  • Identification of oxidation sites
  • O8G oxidation annotation
  • Transcriptome-wide profiling of O8G oxidization
  • Differential oxidation site analysis
  • Length distribution and motif analysis of oxidation sites
  • Evolutionary conservation analysis
  • Explore new O8G oxidation sites
  • GO and Pathway analysis results of differential oxidation sites

Sample Requirements

RNA sample quantity ≥ 5 ug); RNA purity: OD260/280 = 1.8~2.2; OD260/230 ≥ 1.5; RNA quality: 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. Colpaert RM, Calore M. Epigenetics and microRNAs in cardiovascular diseases. Genomics. 2021 Jan 20.
  2. Seok H, Lee H, Lee S, et al. D. Position-specific oxidation of miR-1 encodes cardiac hypertrophy. Nature. 2020 Aug;584(7820).
* For Research Use Only. Not for use in diagnostic procedures.


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