Small RNA Sequencing

Small RNA Sequencing

Service Description

Small RNAs are a type of non-coding RNA (ncRNA) molecules that are less than 200nt in length. They are often involved in gene silencing and post- transcriptional regulation of gene expression. Small RNA sequencing is used to discover novel small RNAs, examine the dierential expression of all small RNAs and to characterize variations with single-base resolution.

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Unique Molecular Identifiers (UMIs) Service Option

Unique Molecular Identifiers (UMIs) can be utilized to eliminate undesirable PCR duplicates derived from a single molecule. After PCR, molecules sharing a UMI are assumed to be derived from the same input molecule. As such, UMI counts offer superior results to counting reads, leading to more accurate estimates of quantitative small RNA expression[1]. UMI technology is especially beneficial to customers undertaking research on rare and precious samples or samples containing less RNAs, such as exosomes.

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DNBseq™ Sequencing Technology

DNBseq™ is an innovative high-throughput sequencing solution, developed by BGI’s Complete Genomics subsidiary in Silicon Valley. The system is powered by combinatorial Probe-Anchor Synthesis (cPAS), linear isothermal Rolling-Circle Replication and DNA Nanoballs (DNB™) technology, followed by high-resolution digital imaging[2].

DNBseq BGI sequencing technology

The combination of linear amplification and DNB technology reduces the error rate while enhancing the signal. The size of the DNB is controlled in such a way that only one DNB is bound per active site on the flow cell. This densely patterned array technology provides optimal sequencing accuracy and increases flow cell utilization.

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Technical Reproducibility

Our DNBseq™ Small RNA sequencing service with optional UMI technology delivers accurate, affordable and high-quality sequencing data to support your academic and clinical research applications.

To demonstrate the high technical reproducibility of the DNBseq™ technology platform for small RNA sequencing, six human brain samples, two heart samples and two blood samples were sequenced [3]. Reproducibility was assessed by using six technical replicates of human brain sample. The median correlation between the six replicates was 0.98, and the 25% and 75% quantile were 0.98 and 0.99, respectively.

Read the Data in our Service Overview

[1]  Fu Y, Wu PH, Beane T, Zamore PD, Weng Z: Elimination of PCR duplicates in RNA-seq and small RNA-seq using unique molecular identifiers. BMC Genomics 2018;19:531.

[2]  Drmanac R et al. : Human genome sequencing using unchained base reads on self-assembling DNA nanoarrays. Science 2010;327:78-81.

[3]  Fehlmann T, Reinheimer S, Geng C, Su X, Drmanac S, Alexeev A, Zhang C, Backes C, Ludwig N, Hart M, An D, Zhu Z, Xu C, Chen A, Ni M, Liu J, Li Y, Poulter M, Li Y, Stahler C, Drmanac R, Xu X, Meese E, Keller A: cPAS-based sequencing on the BGISEQ-500 to explore small non-coding RNAs. Clin Epigenetics 2016;8:123.

 

Project Workflow

We care for your samples from the start through to the result reporting. Highly experienced laboratory professionals follow strict quality procedures to ensure the integrity of your results.

  • Sample QC
  • Library QC
  • Sequencing QC
  • Data QC
  • Sample Preparation
  • Library Preparation
  • Sequencing
  • Raw Data Output
  • Bioinformatics Analysis

How to order

Request a Quote

Send us a no obligation request for quote and our dedicated sales team will reply to you within 24 hours.

Create an Online myBGI Account

Create a myBGI account to view all our sample and shipping documentation and to manage your project moving forwards. It's quick, free and easy!

Sequencing Service Specification

BGI Small RNA Sequencing services are executed with BGI's DNBseq™ Technology Platform.

  • Sample Preparation and Services

    Sample Preparation and Services

    • 50bp single-end sequencing reads
    • Standard output 20 Million reads per sample
    • Clean data and bioinformatics analysis are available in standard file formats
    • Standard and customized bioinformatics data analysis are available
    • Cloud-based data storage and delivery system
  • Sequencing Quality Standard

    Sequencing Quality Standard

    • Q20≥90%
    • Adaptor and null rate 10%
    • Small RNA tag rate 20%
    • Clean reads: no less than quantity the contract needed
  • Turn Around Time

    Turn Around Time

    • Typical 25 working days from sample QC acceptance to data delivery
    • Expedited services are available, contact your local BGI specialist for details

Sample Requirements

We can process your human, plant, animal or microbial samples, with the following general requirements:

Sample Type

Amount, Concentration, Minimum Sample Volume

QC Requirements

Regular Samples

Mass: ≥1μg Concentration: ≥50ng/μl
Sample Volume: 15 μl

RIN≥7.5 (plant/fungi) RIN≥8.0 (human/animal)

FFPE RNA

Mass: ≥1μg Concentration: ≥50ng/μl
Sample Volume: 15 μl

RIN≥2.0 DV200≥30%

Small RNA of
Plasma/Serum/Exosome

Mass: ≥20μg Concentration: ≥2ng/μl
Sample Volume: 10 μl

N/A

Data Analysis

In addition to clean data output, BGI offers a range of standard and customized bioinformatics pipelines for your small RNA sequencing project.
Reports and output data files are delivered in industry standard FASTQ, and Excel file formats with publication-ready tables and figures.

STANDARD BIOINFORMATICS ANALYSIS

  • Data filtering

  • Length distribution of small RNAs

  • Analysis of common and specific sequences between two samples

  • Small RNAs distribution across selected genome

  • Identification of rRNAs, tRNAs, snRNAs, snoRNAs, etc.

  • Identification of repeat associated small RNAs

  • Identification of small RNA sequences which could align to exon/intron

  • Identification of known miRNAs by aligning to designated part of miRBase

  • Analysis of the expression pattern of known miRNAs

  • Classification of small RNAs into several categories based on customer preference

  • Prediction of novel miRNAs and their secondary structures by Mireap and miRDeep from unannotated small RNAs

  • Family analysis of known miRNAs

CUSTOMIZED ANALYSIS

Further customization of Bioinformatics analysis to suit your unique project is available: Please contact your BGI technical representative

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