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circRNA sequencing-Illumina

Circular RNA sequencing (circRNA-seq) is to profile and analyze circular RNAs, a class of RNA molecules that form closed loops due to non-canonical splicing events, providing these RNA with increased stability. While some circRNAs have been shown to act as microRNA sponges, sequestering microRNAs and preventing them from regulating their target mRNAs, other circRNAs may interact with proteins, modulate gene expression, or have roles in cellular processes. circRNA expression analysis provides insights into the regulatory roles of these molecules and their significance in various cellular processes, developmental stages, and disease conditions, contributing to a deeper understanding of the complexity of RNA regulation in the context of gene expression.


Service Details

Bioinformatics

Demo Results

Featured Publications

Features

● rRNA depletion followed by directional library preparation, enabling sequencing data that is strand-specific.

● Bioinformatic workflow enables circRNA prediction and expression quantification

 

Service Advantages

●  More comprehensive RNA libraries: we use rRNA depletion instead of linear RNA depletion in our pre-library preparation, ensuring that the sequencing data includes not only circRNA but also mRNA and lncRNA, enabling joint analysis on these datasets

●  Optional analysis of competitive endogenous RNA (ceRNA) networks: providing deeper insights into cellular regulatory mechanisms

●  Extensive Expertise: with a track record of processing over 20,000 samples at BMK, spanning diverse sample types and lncRNA projects, our team brings a wealth of experience to every project.

●  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

Poly A enriched

Illumina PE150

16-20 Gb

Q30≥85%

Sample Requirements:

Nucleotides:

Conc.(ng/μl)

Amount (μg)

Purity

Integrity

≥ 100

≥ 0.5

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Limited or no protein or DNA contamination shown on gel.

For plants: RIN≥6.5;

For animals: RIN≥7.0;

 5.0≥28S/18S≥1.0;

limited or no baseline elevation

● Plants:

   Root, Stem or Petal: 450 mg

   Leaf or Seed: 300 mg

   Fruit: 1.2 g

● Animal:

   Heart or Intestine: 450 mg

   Viscera or Brain: 240 mg

   Muscle: 600 mg

   Bones, Hair or Skin: 1.5g

● Arthropods:

   Insects: 9g

   Crustacea: 450 mg

● Whole blood: 2 tubes

● Cells: 106 cells

● Serum and Plasma: 6 mL

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

Library Preparation

Sequencing

Data analysis

Data analysis

After sale Services

After-sale services


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  • Bioinformatics

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    circRNA prediction: chromosomal distribution

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    Differentially Expressed circRNAs – Volcano plot

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    Differentially Expressed circRNAs – hierarchical clustering

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    Functional enrichment of circRNA’s host genes

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    Explore the research advancements facilitated by BMKGene’ circRNA sequencing services through a curated collection of publications.

     

    Wang, X. et al. (2021) ‘CPSF4 regulates circRNA formation and microRNA mediated gene silencing in hepatocellular carcinoma’, Oncogene 2021 40:25, 40(25), pp. 4338–4351. doi: 10.1038/s41388-021-01867-6.

    Xia, K. et al. (2023) ‘X oo-responsive transcriptome reveals the role of the circular RNA133 in disease resistance by regulating expression of OsARAB in rice’, Phytopathology Research, 5(1), pp. 1–14. doi: 10.1186/S42483-023-00188-8/FIGURES/6.

    Y, H. et al. (2023) ‘CPSF3 modulates the balance of circular and linear transcripts in hepatocellular carcinoma’. doi: 10.21203/RS.3.RS-2418311/V1.

    Zhang, Y. et al. (2023) ‘Comprehensive evaluation of circRNAs in cirrhotic cardiomyopathy before and after liver transplantation’, International Immunopharmacology, 114, p. 109495. doi: 10.1016/J.INTIMP.2022.109495.

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