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Exosomal mRNA/LncRNA/CircRNA Sequencing-Illumina

Exosomes are small vesicles secreted by cells, typically ranging in diameter from 30 to 100 nanometers. These vesicles contain various RNA. Exosomes are believed to play crucial roles in intercellular communication, immune responses, and disease development, and can be disseminated to other parts of the body through bodily fluids such as plasma, saliva, and urine. They carry specific biomolecules to regulate the functions of recipient cells, influencing cellular physiological states. Exosomes are also considered to play key roles in disease development, including cancers, neurodegenerative diseases, and inflammatory conditions. Research on exosomes offers new insights and methods for the diagnosis, treatment, and prevention of diseases.


Service Details

Bioinformatics

Demo Results

Features

● Special probes are used to deplete the ribosomal cDNA.

● Strand-specific library construction.

● One workflow to get mRNA, lncRNA and circRNA data simultaneously.

Service Advantages

● Rigorous Quality Control: We implement core control points across all stages, from sample and library preparation to sequencing and bioinformatics. This meticulous monitoring ensures the delivery of consistently high-quality results.

● Post-Sales Support: Our commitment extends beyond project completion with a 3-month after-sale service period. During this time, we offer project follow-up, troubleshooting assistance, and Q&A sessions to address any queries related to the results.

Sample Requirements and Delivery

Library

Platform

Recommended data

Data QC

rRNA depleted directional library

Illumina PE150

10-16 Gb

Q30≥85%

Sample Requirements

Nucleotides:

 Amount (ng)

Volume (μL)

40

 20

Tissue:

Exosome: 100 μL; 40 μg; >10^9 particles/ mL
Serum/plasma: 5 mL
Cell culture supernatant: 100 mL
Urine: 100 mL
Saliva: 15 mL
Nasal discharge: 15 mL
Cerebrospinal fluid: 15 mL
Amniotic fluid: 50 mL
Semen: 5 mL
Bile: 5 mL
Ascites: 50 mL
Milk: 15 mL
Follicular fluid: 15 mL
Fecal supernatant: 50 mL
Microbial culture supernatant: 100 mL
Animal tissue: 3 g
Plant tissue: 1 g

Recommended Sample Delivery

Container: 2 mL centrifuge tube (Tin foil is not recommended)
Sample labeling: Group+replicate e.g. A1, A2, A3; B1, B2, B3.
Shipment:
1. Dry-ice: Samples need to be packed in bags and buried in dry-ice.
2. RNAstable tubes: RNA samples can be dried in RNA stabilization tube(e.g. RNAstable®) and shipped in room temperature.

Service Work Flow

Sample QC

Experiment design

sample delivery

Sample delivery

Pilot experiment

RNA extraction

Library Preparation

Library construction

Sequencing

Sequencing

Data analysis

Data analysis

After sale Services

After-sale services


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  • Bioinformatics

    BioinformaticsBioinformatics2 

    circRNA prediction: chromosomal distribution

     图片36

     

    Differentially Expressed circRNAs – Volcano plot

     图片37

     

    Differentially Expressed circRNAs – hierarchical clustering

     图片38

     

    Functional enrichment of circRNA’s host genes

     图片39

     

    Differential Gene Expression (DEGs) analysis

     

     图片30

     

     

    Quantification of lncRNA expression – clustering

     

    图片31 

     

    Enrichment of lncRNA target genes

     

     图片32

     

    Joint mRNA and lncRNA position analysis – Circos plot (middle circle is mRNA and inner cicrlce is lncRNA)

     

     图片33

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