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CircRNA Sequencing

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By supporting the recent emerging of circular RNAs research (circRNAs), CD Genomics is providing circRNA sequencing service through deep sequencing to meet your project requirement and budget. We offer a complete service including experimental design, RNA extraction, library preparation, sequencing, and bioinformatics analysis.

CircRNA Sequencing

CircRNAs are a recent novel class of abundant, stable and ubiquitous RNAs addition to the growing list of types of non-coding RNA molecules, which were first identified in the early 1990s. CircRNAs have previously evaded recognition or simply been discarded as artefacts during standard processing of high-throughput RNA-seq data. CircRNAs are pervasively expressed RNA molecules in which a covalent and canonical linkage termed a “backsplice” has formed between a downstream 3’ splice site and an upstream 5’ splice site in a linear pre-messenger RNA. CircRNAs are distinguished from mRNAs in that they lack poly (A) tails and 5ʹ caps, and the abundance of circRNA is approximately 2–4% of the total mRNA in cells. CircRNAs can arise from both exons (exonic circRNA) and introns (intronic circRNA). CircRNAs can serve as miRNA or RNA-binding protein ‘sponges’, sequestering miRNAs and preventing their interactions with target mRNAs, as a result, controlling transcriptional events. Study of this class of non-coding RNAs has potential implications for therapeutic and research applications. Comprehensive detection of circRNAs from high-throughput transcriptome data is an initial and crucial step to study their biogenesis and function.

CircRNA Sequencing

The high-throughput sequencing of rRNA-depleted RNA, in combination with computational tools, has led to the identification of thousands of new circRNAs and analysis of their linear host transcripts in a quantitative manner in diverse organisms.

Circular RNAs, unlike miRNAs and other small RNAs, are not easily separated from other RNA species by size or electrophoretic mobility. CircRNAs are resistant to exonuclease treatment, whereas linear RNAs are selectively depleted with this approach. Consequently, circRNAs can be retained in rRNA-depleted libraries and enriched in libraries treated with RNase R to digest linear RNA. Identification and characterization of circRNA transcripts was done computationally with bioinformatics tools such as CIRI. Relative abundance changes in circRNA expression are typically approximated by comparing normalized back-splice junction read counts of specific circRNAs under different conditions and/or time points.

Data Analysis

Our data analysis programs following circRNA sequencing includes circRNA identification and annotation, known circRNA expression analysis (differential expression analysis, enrichment analysis and functional analysis), novel circRNA prediction, origin analysis of circRNA, and target miRNA prediction.

Sequencing Strategy and Recommended Depth

  • Illumina HiSeq PE150
  • ≥ 10 Gbase

Sample Requirements

  1. Sample type: Total RNA without degradation or DNA contamination.
  2. Starting amount of total RNA: ≥ 10 µg
  3. Sample conc.: ≥ 100 ng/µl
  4. Sample purity: OD260/280 = 1.8~2.2

Key Features and Advantages

  • Predict circRNAs targets: We can predict the target which plays a regulation role by the information analysis.
  • Low false positive: Obtain the accurate and reliable circRNA information as per the analysis methods by eliminating the interference of linear RNA.
  • Cost-effective: Our high success rates and quality data prevent costly repeat experiment and sequencing.
  • Highly experienced personnel: CD genomics helps you to design your project and offers consultation with you at every stage of the process.

Supported by our experienced scientists and advanced technology, CD genomics can help you acquiring the circRNA sequence information with single-base resolution in one time through the high-throughput sequencing by following the construction of strand-specific library for the simultaneous removal of rRNA and linear RNA, and identify the known circRNA and predict the novel circRNA and its target gene based on powerful bioinformatics analysis platform.

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