RNA Bisulfite Sequencing Service

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RNA Bisulfite Sequencing Service

IntegrateRNA offers a whole transcriptome-wide, base resolution RNA methylation (m5C) analysis service using advanced sequencing technique. We utilize the classic bisulfite processing and next-generation sequencing platforms to detect m5C methylation modification on whole transcriptome range. We are dedicated to providing a high-resolution, efficiency and economy approach for m5C detection and localization in various RNA molecular, such as mRNA, tRNA, rRNA.

The nucleobase modification 5-methylcytosine (m5C) is widespread both in DNA and different cellular RNAs. 5-methylcyosine is emerging as an important epitranscriptomic mark of RNA. Various RNA methyltransferases that reside at different locations in the cell install this mark on a variety of RNA types. These range from cytoplasmic and mitochondrial rRNA and tRNA to mRNA and to various other non-coding RNA. Similar to the multitude of targets, many if not most aspects of RNA metabolism may be affected by the m5C mark. Recent data strongly suggest that RNA cytosine methylation affects the regulation of various biological processes including RNA stability and mRNA translation. In addition, m5C labeling is a versatile tool for fine-tuning RNA processing, stability, translation, and RNA-protein interactions.

The study of methylation at single base resolution of individual cytosines in RNA is facilitated by bisulfite treatment of RNA sample followed by PCR amplification, cloning, and sequencing of individual amplimers. (Figure1)

Schematic diagram of m5C detection by RNA bisulfite sequencingFigure 1. Schematic diagram of m5C detection by RNA bisulfite sequencing

Workflow of RNA Bisulfite Sequencing

  • Total RNA extraction and quality control
  • Bisulfite treatment (Bisulphite conversion)
  • RNA purification
  • Library preparation and high-throughput sequencing (Bisulfite treated and untreated input sample)

Data Analysis Pipeline

RNA Bisulfite Sequencing Service

Data Analysis Items

  • m5C modification level analysis
  • Statistic and annotation of m5C modification site
  • Identification and statistic of differentially methylated cytosine
  • Custom-tailored data analysis

Sample Requirement

  • Total RNA: >100ug (Conc: >100ng/ul; RIN≥7; Ratio 28S/18S ≥0.7)
  • Cell: >10*8
  • Tissue: >300mg (Cut large tissue samples to ≤ 0.2cm in any single dimension. Plate three pieces of tissue in 1ml RNAlater solution.)

IntegrateRNA's next-generation sequencing platforms can deliver a great amount of useful RNA methylation information with publication-ready data parsed by our expert bioinformatics scientists.

Reference:

  1. Yuri, M. et al. (2009). 5-methylcytosine in rna: detection, enzymatic formation and biological functions. Nucleic Acids Research, 38(5), 1415-1430.
For research use only. Not intended for any clinical use.
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