Small RNA Sequencing

Small RNAs are short (<200 nt in length) and usually non-coding RNA molecules. There are several biologically relevant and functionally diverse classes of sRNA of specific sizes and produced by different, genetically separable pathways. Small RNAs include miRNA, siRNA, piRNA, snoRNA, snRNA, tsRNA, and srRNA. One of the most representative and significant functions of small RNA is RNA silencing (RNAi), in this case, endogenously expressed microRNA (miRNA) or exogenously derived small interfering RNA (siRNA) induces degradation of the complementary mRNA.

Overview

The innovation of large-scale, next-generation sequencing has exponentially increased knowledge of RNA biology, with regard to the diversity, abundance, and function of various RNA molecules. Small RNA-seq is a powerful tool for analyzing small RNAs such as miRNAs, siRNAs, and piRNAs in a single sequencing run, allowing the evaluation and discovery of novel small RNA molecules and the prediction of their functions. These RNA-seq methods have provided an even more complete characterization of small RNA and promised further applications.

Our technologies at single-base resolution allow for small RNA detection from very small amounts of cellular materials, which can help you detect pre-known small RNA, discover new small RNA, and examine all small RNA for differential expression in any sample. We generate small RNA sequencing libraries directly from total RNA and Capture the complete range of small RNAs, to understand the roles they play. this will provide you with a comprehensive and efficient approach to understand post-transcriptional regulation and to discover novel biomarkers.

Service Portfolio

Explore how biofluid profiling using NGS helps researchers understand functions and dynamics of small RNA.

UMI Small RNA Sequencing

Our small RNA sequencing combined with UMI technology recognizes PCR duplicates derived from a single molecule and delivers accurate, affordable sequencing data, leading to accurate estimates of quantitative small RNA expression.

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Bacterial Small RNA-Seq

Bacterial small RNA sequencing enables the detection of novel bacterial small RNA molecules, profiling of small RNA expression, prediction of small RNA targets, hence facilitating function and mechanism studies.

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miRNA Sequencing

miRNA sequencing profiles microRNAs in any species, enabling the discovery of novel miRNAs and revelation of the importance of miRNAs linked to a wide spectrum of biological functions.

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piRNA Sequencing

piRNAs are closely associated with the development of germ cells and axoneuron. We provide piRNA sequencing to capture all piRNAs in a sample, allowing for piRNA screening and research.

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Features

Superb Expertise Transcriptome-Wide Accurate Annotation One-Stop Solution
Decade of experience in high-throughput sequencing and bioinformatics analysis. Profile various small RNA, including miRNA, siRNA, and piRNA. Provide complete annotate information with multiple databases for small RNAs. One-stop solution from sample QC and 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

RNA fragmentation;
m7G/m3C 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.

Small RNA Sequencing

Bioinformatics Analysis Pipeline

In-depth data analysis:

  • Length distribution and quantitative analysis of small RNA
  • Differential expression analysis of small RNA
  • Prediction of small RNA
  • Discovery of novel biomarkers
  • Coding-non-coding gene co-expression network (CNC network)
  • GO annotation analysis
  • KEGG pathway analysis
  • Interactive network analysis

Sample Requirements

RNA sample (concentration ≥ 1 ng/uL, quantity ≥ 2 ug)

1.8 ≤ OD260/280 ≤ 2.2, OD260/230≥2.0, RIN ≥ 6.5, 28S:18S ≥ 1.0.

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. McCormick KP, Willmann MR, Meyers BC. Experimental design, preprocessing, normalization and differential expression analysis of small RNA sequencing experiments. Silence. 2011, 2(1):1-9.
  2. Sharma CM, Vogel J. Experimental approaches for the discovery and characterization of regulatory small RNA. Current opinion in microbiology. 2009, 12(5):536-46.
  3. Burroughs AM, Ando Y, Aravind L. New perspectives on the diversification of the RNA interference system: insights from comparative genomics and small RNA sequencing. Wiley Interdisciplinary Reviews: RNA. 2014, 5(2):141-81.
* For Research Use Only. Not for use in diagnostic procedures.


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