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LncRNA Sequencing

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CD Genomics is offering high-throughput and cost-efficient lncRNA sequencing service by combining the latest Illumina sequencing instruments and advanced bioinformatics analysis.

The Introduction of LncRNA Sequencing

Long non-coding RNAs (lncRNAs) are defined as a large and diverse class of transcribed RNAs with size greater than 200 nt that do not encode proteins. LncRNAs are widely distributed in organisms and lncRNA transcripts account for the major part of the non-coding transcriptome. LncRNAs may be classified into different subtypes (including antisenses, intergenic, overlapping, intronic, bidirectional, and processed) based on the position and direction of transcription in relation to other genes. LncRNAs resemble mRNAs because they are typically transcribed from active chromatin, polyadenylated, and capped. However, they do not direct protein synthesis.

LncRNAs are functionally important to organisms and not merely the product of transcriptional noise. A myriad of molecular functions of lncRNAs have been discovered in mammals and plants, including nucleosome repositioning, chromatin remodeling, transcriptional control, and posttranscriptional processing. LncRNAs are increasingly implicated in disease occurrence, genomic imprinting and developmental regulation. Gene expression profiling and in situ hybridization studies have revealed that lncRNA expression is developmentally regulated, can be tissue- and cell-type specific, and can vary spatially, temporally, or be in response to stimuli.

The application of next-generation sequencing technology has greatly facilitated the discovery and function analysis of lncRNAs. LncRNA sequence information can be acquired at single-base resolution via library preparation, high-throughput sequencing, and powerful bioinformatics analysis. We construct the sequencing library by the removal of rRNA and retain both lncRNAs and mRNAs. The lncRNA-mRNA interaction analysis contributes to the illumination of lncRNA regulatory networks.

Advantages of LncRNA Sequencing

  • Identifies known and novel features
  • Allows profiling of lncRNAs across a wide dynamic ranges
  • Explores novel biomarkers and lncRNAs regulatory networks

LncRNA Sequencing Workflow

The general workflow for lncRNA sequencing is outlined below. To construct lncRNA sequencing library, the first step of lncRNA sequencing is to deplete rRNA, followed by RNA fragmentation, cDNA synthesis, adaptor ligation, size selection and PCR enrichment. Our highly experienced expert team executes quality management, following every procedure to ensure confident and unbiased results.

Service Specification

Sample requirements and preparation
  • Sample type: Total RNA without degradation or DNA contamination
  • Starting amount of total RNA ≥ 2 µg
  • Sample concentration ≥ 200 ng/µl
  • Sample purity: OD260/280 = 1.8~2.2
  • All RNA samples are validated for purity and quantity
Sequencing
  • Strand-specific library preparation
  • Illumina HiSeq PE150
  • ≥10G data for small genome and ≥12G for large genome
  • More than 80% of bases with a ≥Q30 quality score
Bioinformatics Analysis
We provide customized bioinformatics analysis including:
  • Raw data quality control and length filter
  • Reference-based mapping
  • Prediction of novel transcripts (including lncRNAs)
  • Quantification and differential expression analysis of lncRNAs and mRNAs
  • Classification of lncRNA family and lncRNA functional annotation
  • SNP/InDel calling, identification of splicing variants
  • LncRNA target prediction and lncRNA-mRNA interaction analysis

Analysis pipeline

Our PhD-level bioinformatics team provides comprehensive analysis for both lncRNAs and mRNAs, enabling access to lncRNA and mRNA information in a single sequencing run. We can help in the experimental design at the very beginning of your project and offer consultation at every stage of the project process.

Related Services
RNA-Seq

Transcriptome mapping and quantification

Small RNA Sequencing

Expression profiling of small RNAs

CircRNA Sequencing

Expression profiling of circular RNAs

Degradome Sequencing

Insight into microRNAs landscape

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CONTACT CD GENOMICS

45-1 Ramsey Road, Shirley, NY 11967, USA
Tel: 1-631-275-3058
Fax: 1-631-614-7828
Email: info@cd-genomics.com