Biofluid Small RNA-Seq

Biofluid small RNA sequencing is a method that combines the next-generation sequencing (NGS) with optimized RNA handling and library preparation. It can improve the quality of small RNA sequencing in the biofluid and can accurately and comprehensively analyze small RNA molecules, including siRNAs, piRNAs, snoRNAs, snRNAs at one time. It is highly recommended for the analysis of small RNA variations and rare sequences.

Overview

Small RNAs, which are defined as short (>200 nucleotides, often 18 to 30 nucleotides), usually non-coding RNA molecules, consist of a diverse category of regulatory RNAs including microRNA (miRNA), small interfering RNA (siRNA), piwi-interacting RNA (piRNA), small nucleolar RNA (snoRNA), small nuclear RNA (U-RNA or snRNA), tRNA-derived small RNA, etc. Small RNA sequencing (RNA-seq) approach allows us to profile small RNA molecules, discover new forms of small RNA, detect differentially expressed RNAs and predict their possible functions. Biofluid small RNA sequencing provides credible data with professional bioinformatics analysis, which can truly reflect the abundance of transcript expression, and achieve unbiased and accurate quantification for small RNA in biofluid samples such as exosomes. Biofluid small RNA sequencing achieves high precision in base calling and quantification for small RNAs from biofluid in a specific space-time state. With optimized RNA handling and library preparation, our small biofluid RNA-seq service can facilitate studies with a variety of human biofluid samples. As circulating small RNAs are found associated with a range of human diseases, biofluid small RNA sequencing can be used to study human diseases such as cancer, myocardial injuries, hepatic diseases, and neurodegenerative diseases and identify potential biomarkers.

Features

Various Samples Transcriptome-Wide Bioinformatics Analysis One-Stop Solution
This method can be applied to a variety of biofluid samples including exosomes. Flexibility to focus sequencing on multiple small RNA classes including miRNA, tRNA, piRNA, and snoRNA. Custom bioinformatics analysis that focuses on specific small RNA classes of interest to satisfy customer needs. One-stop solution from sample QC, library construction, to sequencing and data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

QC and quantification;
rRNA removal.

Library Preparation

2. Library Preparation

Size selection (17 to 35 nt or a custom range), cDNA library construction, library QC by BioAnalyzer and RTQ-PCR.

Sequencing

3. Sequencing

NextSeq500/HiSeq2500;
50/75/100 nt, single or paired-end.

Data Analysis

4. Data Analysis

Visualize and preprocess results, and perform custom bioinformatics analysis.

Bioinformatics Analysis Pipeline

Biofluid Small RNA-Seq

In-depth data analysis:

  • Profiling of miRNA, siRNA, piRNA, snoRNA, snRNA
  • Predict novel miRNA
  • Detect novel and rare transcript variants
  • Differential expression analysis of transcripts
  • Target gene prediction and functional analysis of siRNA, piRNA, snoRNA, snRNA, and miRNA
  • GO enrichment analysis
  • KEGG pathway analysis
  • Annotation of miRNA, siRNA, piRNA, snoRNA, snRNA

Sample Requirements

RNA sample (concentration ≥ 1 ng/uL, quantity ≥ 20 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, full statistical analysis & alignments, custom analysis reports on customer request.

References:

  1. Srinivasan S, Yeri A,Cheah P S, et al. Small RNA Sequencing across Diverse Biofluids Identifies Optimal Methods for exRNA Isolation. Cell, 2019, 177: 446-462.
  2. Godoy P M, Bhakta N R,Barczak A J, et al. Large Differences in Small RNA CompositionBetween Human Biofluids. Cell Reports, 2018,25: 1346-1358.
  3. 3.Yeri A, Courtright A, Reiman R, et al. Total Extracellular Small RNAProfles from Plasma, Saliva, and Urine of Healthy Subjects. Scientific REPORTS, 2016, 7:44061.
* For Research Use Only. Not for use in diagnostic procedures.


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