全トランスクリプトームシーケンス

概要
デモ結果

概要

Novogene’s Whole Transcriptome Sequencing service equips the research with cutting-edge NGS solutions that provide in-depth bioinformatic analysis on all transcripts including mRNAs and non-coding RNAs. This competitive approach investigates and explores potential transcriptional and regulatory network mechanisms, while providing key insights into interaction and intersection functionality from a comprehensive transcriptomic perspective.

Service Specifications

Applications

Co-localization and co-expression of ncRNA and mRNA detection
miRNA sponge and target regulatory elements detection
Regulatory network investigation: ceRNA regulatory network set up based on lncRNA/circRNA-miRNA-gene pairs, taking lncRNA/circRNA as decoy, miRNA as core and mRNA as target

Advantages

Extensive experience with thousands of samples being successfully sequenced.
Unsurpassed data quality with a guaranteed Q30 score ≥ 80% that exceeds Illumina’s official benchmarks.
Comprehensive analysis using mainstream software and mature in-house pipeline to meet multiple bioinformatic requests.

Sample Requirements

Library Type	Sample Type	Amount	RNA Integrity Number (Agilent 2100)	Purity (NanoPore)
lncRNA Library & small RNA Library	Total RNA	≥ 5 μg	Animal ≥ 7.5, Plant ≥ 7, with smooth baseline	OD260/280 = 1.8-2.2; OD260/230 ≥ 1.8;

Sequencing Parameter and Analysis

Platform	Illumina Novaseq 6000
Read length	Paired-end 150 & Single-end 50
Recommended Sequencing Depth	≥40 million read pair per sample (lncRNA library); ≥20 million read pair per sample (small RNA library);
Association Analysis of Transcriptome	Interaction of lncRNA and miRNA
	Interaction of mRNA and miRNA
	Interaction of circRNA and miRNA
	Regulatory Network of lncRNA, miRNA and mRNA
	Regulatory Network of circRNA, miRNA and mRNA

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Project Workflow

Sampling:

Total RNA was extracted from the spinal cord dorsal horn tissue

Sequencing Strategy:

1. lncRNA library: NEBNext UltraTM Directional RNA Library Prep Kit for Illumina
2. small RNA library: NEBNext Multiplex Small RNA Library Prep Set for Illumina
3. sequenced on Illumina HiSeq 2500 platform, 125 bp paired-end and 50-bp single-end reads, respectively.

Figure 1. The expression profiling changes of lncRNAs in spinal cord of SNI rats Vocalno Plot indicate up and down regulated lncRNAs of rats in group SNI compared with group CON (A)

Figure 2. The expression profiling changes of circRNAs in spinal cord of SNI rats Vocalno Plot indicate up and down regulated circRNAs of rats in group SNI compared with group CON (A)

Figure 3. The expression profiling changes of miRNAs in spinal cord of SNI rats Vocalno Plot indicate up and down regulated miRNAs of rats in group SNI compared with group CON (A)

Figure 4. The expression profiling changes of mRNAs in spinal cord of SNI rats Vocalno Plot indicate up and down regulated mRNAs of rats in group SNI compared with group CON (A)

Figure 5. Counts of relatived ncRNAs and mRNAs in spinal cord of SNI rats.

Figure 6. lncRNA-micRNA-mRNAs regulatory network analysis of ncRNAs in spinal cord of SNI rats.

Figure 7. cirRNA-micRNA-mRNAs regulatory network analysis of ncRNAs in spinal cord of SNI rats.

Conclusion:

This study comprehensively identifies regulated ncRNAs of the spinal cord and to demonstrate the involvement of different ncRNA expression patterns in the spinal cord of NP pathogenesis by sequence analysis. This information will enable further research on the pathogenesis of NP and facilitate the development of novel NP therapeutics targeting ncRNAs.