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RNA-seq services

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  •  Description
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Catalog Number
Format
G02030000

Diagenode uses proprietary ligation-free CATS-based technology for RNA-seq library preparation

Read more about CATS

  • Ideal for low input samples including patient samples
  • Proven to detect more transcripts than any other commercial method
  • Full range of RNA-seq services including the quantification of known transcripts:
    • small RNA-seq (including small non-coding RNA like miRNA)
    • mRNA-seq
    • Long non-coding RNA-seq
    • Whole transcriptome sequencing

Excellent performance on challenging samples

Heatmap clustering of the top 50 differentially expressed microRNAs(miRNAs) after library preparation:

  • CATS Small RNA-seq Kit on 1 ng exosomal RNA (Exosome sample 1, 2)
  • CATS RNA-seq Kit on 1 ng total RNA from parent (HEK293T) cells (Cell sample 1, 2).

Prior library preparation, exosomes were isolated with Diagenode exosome capture solutions Exosomes

Unique miRNA (>1 copy) detected in circulating plasma RNA

Transcripts detected in miRNA samples of CATS small RNA-seq Kit vs Competitor Kit.
TPM >1 and TPM>10

Input: 1 ng circulating small RNA in plasma samples.
PCR cycles: CATS 14x vs Competitor 21x. Final library yield: CATS 27 ng vs Competitor 1.2 ng.

More miRNA transcripts detected from challenging samples

Transcripts detected in miRNA samples of CATS small RNA-seq Kit vs Competitor Kit.

Input: 1 ng circulating small RNA in plasma samples.
PCR cycles: CATS 14x vs Competitor 21x. Final library yield: CATS 27 ng vs Competitor 1.2 ng.

Ultra low inputs, few PCR cycles

Representation of the library preparation yield according to different methods (CATS small RNA-seq Kit vs NEBNext UltraI kit) for 100 ng total RNA. Number of PCR cycles applied following manufacturer’s instructions.

Consistency across template switching techniques

Comparison of the number of detected small non-coding RNAs CATS small RNA-seq Kit vs SMARTer
Input: 100 pg isolated small RNA. TPM ≥2. Transcripts taken into consideration: miRNA, piRNA, scRNA, scaRNA, snoRNA, snRNA and vaultRNA. 1,217 transcripts found in both libraries at this expression level.

CATS saves time

Very reproducible library preparation method

Transcripts detected in two technical replicates of CATS v2 total RNA-seq libraries. TPM≥2
Input: 1 ng rRNA(-) RNA from human universal total RNA (Agilent, 740000).

Broad overview of the transcriptome

Biotypes* represented in the libraries CATS v2 mRNA-seq Kit vs NEBNext Ultra I directional RNA-seq Kit. TPM ≥2.
Input: 1 ng, 10 ng poly(A) RNA from human universal total RNA (Agilent, 740000). * < 0.04% of total detected transcripts at this threshold is rRNA

More transcripts detected

Biotypes* represented in the libraries CATS v2 total RNA-seq Kit vs NEBNext Ultra I directional RNA-seq Kit. TPM ≥2.
Input: 1 ng, 10 ng rRNA(-) RNA from human universal total RNA (Agilent, 740000). * < 2% of total detected transcripts at this threshold is rRNA

Uniquely detected transcripts

All detected transcripts in CATS v2 mRNA-seq Kit vs NEBNext Ultra I directional library preparation Kit. No threshold has been set for the analysis.
Input: 1 ng poly(A) RNA from human universal total RNA (Agilent, 740000). 36,245 transcripts found in both libraries.

  • Description
    RNA quality control
    • Measurement of concentration as well as determination of RIN value (RNA integration number).
    Library preparation
    • CATS RNA-seq library preparation, a ligation-free proprietary technology that retains strand specificity is used.
    • Positive control RNA and a negative control (H2O) are included in parallel. Upon request a spike-in (ERCC spike-in mix or cel-miR-39-3p) can be also included.
    • Final QC prior to sequencing (quantification and fragment size).
    Sequencing
    • Single-read, 50 bp (paired-end on request)
    • miRNA and small RNA-seq: on average 10 million reads/sample.
    • mRNA-seq: on average 40 million reads/sample.
    • Total RNA-seq: on average 40 million reads/sample.
  • Bioinformatic analysis

    Analysis

    Features
    Standard Alignment to reference genome and quantification of known genes and/or transcripts is included.
    Differential expression differential expression testing between samples for genes and/or transcripts.
    Gene ontology terms analysis analysis of differentially expressed gene sets. Gene Ontology terms that are overrepresented in differentially expressed genes may indicate the underlying biological processes involved
    Pathway analysis identification of biochemical pathways in which genes associated with differential expression may be overrepresented.

  •  Applications
    Small RNA and miRNA
    Post-transcriptional regulation of gene expressionNovel transcripts and differentially-expressed small RNAsmiRNAs involved in regulation of onco- and tumor-suppressor gene expressionDifferential miRNA expression Read more
    Messenger RNA (mRNA)
    Protein-coding RNA with specific poly A selectionCancer-related mRNA signaturesAberrant mRNA translation in cancer pathogenesisGene expression profiling and identification of differentially expressed genes (DEG)Epigenetic mechanism of gene express... Read more
    Whole transcriptome analysis and long non-coding RNA (lncRNA) analysis
    Transcriptome sequencing: Analysis of whole transcriptome and RNAs larger than 200nt with optional rRNA depletionNon-protein coding RNAs involved in chromatin remodeling & transcriptional/post-transcriptional regulation Read more
  •  Documents
    The Diagenode Epigenetics custom service POSTER
    Complete workflows for genome-scale DNA methylation and histone marks analysis Epigenetics is cr...    		 Download
  •  Publications

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