Circular RNA sequencing (circRNA-seq) profiles and analyzes circular RNAs, a class of RNA molecules that form closed loops due to non-canonical splicing events, providing this 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.

Platform: Illumina Novaseq X

  Features

- rRNA depletion followed by directional library preparation, enabling strand-specific sequencing data.
- Bioinformatic workflow enables circRNA prediction and expression quantification.

Service Advantages 

- Extensive Expertise: We have processed over 20,000 samples, spanning diverse sample types, we bring the wealth of expertise to every project.
- Rigorous Quality Control: We implement core control points across all stages, from sample preparation to library preparation, sequencing and bioinformatics. Our meticulous monitoring ensures the delivery of consistently high-quality results.
- 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 of these datasets.
- Optional Analysis of Competitive Endogenous RNA (ceRNA) Networks: Providing deeper insights into cellular regulatory mechanisms.
- Post-Sales Support: We understand how important is to be present, that’s why 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

  Sample Requirements:

Nucleotides:

- 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 tube

- 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

Experiment design	Sample delivery	RNA extraction	Library construction	Sequencing	Data analysis	After-sale services

- Data QC
- Genome alignment
- CircRNA identification
- CircRNA expression
- Differential expression analysis

circRNA prediction: chromosomal distribution

Differentially Expressed circRNAs – Volcano plot

Differentially Expressed circRNAs – hierarchical clustering

Functional enrichment of circRNA’s host genes

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.