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CATS RNA-seq Kit v2 x24

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Catalog Number
Format
Price
C05010041
24 rxns
$1,410.00
Other format
C05010045
12 rxns
C05010049
96 rxns

Diagenode’s CATS RNA-seq Kit utilizes the innovative “Capture and Amplification by Tailing and Switching” (CATS), a ligation-free method to produce DNA libraries for next generation sequencing from low input amounts of RNA.

  • Diverse transcripts detection
  • Excellent reproducibility with ultra low inputs down to 100 pg
  • Great performance on challenging samples
  • Read more

    CATS relies on polynucleotide tailing for capturing the 3’-ends of nucleic acids and the template switching ability of MMLV-RT to capture the 5’ ends for cDNA synthesis. This kit allows for generating Illumina compatible ready-to-sequence libraries from RNA inputs as low as 100 picograms (rRNA depleted or polyA selected) in just a few hours.

    Libraries retain strand specificity of origin. Unlike competing solutions, our CATS RNA-seq kit exhibits higher library efficiency versus ligation-based methods providing higher complexity from better template capture and minimal bias due to lower amplification requirements.

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.

Consistency and wide representation of the transcriptome

Comparison on the number of coding transcripts detected with CATS v2 mRNA-seq Kit vs SMARTer stranded RNA-seq library preparation kit. TPM ≥2.
Input: 1 ng poly(A) RNA from human universal total RNA (Agilent, 740000). 21,333 transcripts found in both libraries.

High-complexity at low inputs

Mapped reads distribution for CATS v2 mRNA-seq Kit vs NEBNext Ultra I directional RNA-seq Kit.
Input: 1 ng, 10 ng poly(A) RNA from human universal total RNA (Agilent, 740000).

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).

High correlation in transcripts detection across ultra low inputs

Transcripts detected in 1 ng vs 0.1 ng samples of CATS v2 mRNA-seq libraries. TPM≥2.
Input: 1 ng, 0.1 ng poly(A) RNA from human universal total RNA (Agilent, 740000).

High correlation in coding transcripts detection across different inputs

Transcripts detected in 10 ng vs 1 ng samples of CATS v2 total RNA-seq libraries. TPM≥2
Input: 1 ng, 10 ng rRNA(-) RNA from human universal total RNA (Agilent, 740000).

Few PCR cycles, less amplification bias

BioAnalyzer® electropherogram of two technical replicates of CATS v2 total RNA-seq libraries.
Input: 1 ng rRNA(-) RNA from human universal total RNA (Agilent, 740000).

Excellent performance on challenging samples

Heatmap clustering of the top 50 differentially expressed 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

CATS saves time

  • Characteristics
    • Ligation-free  assay - increased efficiency and minimum bias
    • Low inputs down to 100 picograms
    • User-friendly: reduced hands-on  time with fewer steps than competing methods
    • Libraries ready to sequence in only 5 hours
    • Retaining strand specificity of origin

    CATS RNA-seq library preparation workflow

    Total time Hands-on-time
    (1)

    SAFE STORAGE

    STEP 1
    ssRNA end-repair
    and polyA tailing

    		 75 min 10 min
    (2)

    STEP 2
    RT and template switch

    		 150 min 10 min
    (3)

    SAFE STORAGE

    STEP 3
    PCR amplification
    of DNA library

    		 30 min 5 min
    (4)

    SAFE STORAGE

    (1) RNA: stored at -80°C after purification (safe stop & storage) ; (2) cDNA: stable at 4°C overnight, store at -20°C ; (3) Amplified library: prior purification and QC store at -20°C (safe stop & storage) ; (4) DNA libraries: store at -20°C or -80°C (safe stop & storage)

    Schematic representation of the workflow used by the CATS Small RNA-seq Kit. Single stranded RNAs are first dephosphorylated (end-repaired) and polyadenylated at the 3’-end. Subsequently, a cDNA strand synthesis is performed in the presence of the anchored poly(dT) oligonucleotide containing terminal P7 Illumina® adaptor sequence. When the reverse transcriptase reaches the 5’-end of the RNA it switches the template and continue DNA synthesis over the template-switching oligonucleotide (TSO). The TSO contains three 3’-terminal ribonucleotides X (rX) which facilitate the template switching and carry the terminal P5 Illumina® adaptor sequence. During PCR pre-amplification of the first cDNA strand, Illumina® adapters carrying P5 and P7 terminal sequences (required for clustering on an Illumina® flow cell) as well as index sequences are incorporated into the library. The sum size of the adapters (the size of “empty” library) is 143 bp.

  • Data analysis and interpretation

    Single-click visual informatics and gene expression analysis for CATS RNA-seq kits powered by Genialis™ Expressions software

    Take control of your data with a fully automated data processing pipeline developed and validated for use with Diagenode’s CATS RNA-seq kits. Combined with Genialis’ guided data interpretation workflows in their interactive visual interface, you can:

    • Establish reproducibility across projects. Run your raw data through the CATS-RNA-seq pipeline so your primary data analyses are done the same way across projects.
    • Ensure experiment design quality. Verify consistency between your experimental replicates and identify relevant correlations or differences amongst experimental conditions.
    • Compare expression levels across conditions. Quantify gene abundance in individual samples and investigate differential expression of individual genes, predefined gene sets, and entire transcriptomes.
    • Assess differences in biological activity. Understand how specific experimental conditions affect the regulation of groups of related genes and their corresponding biological functions.

    To learn more about the CATS RNA-seq data analysis workflow, visit genial.is/CATS-workflow.

    See CATS RNA-seq data analysis in action at genial.is/CATS-tutorial-video.

    Request a personal walkthrough of CATS RNA-seq data analysis on Genialis’ Expressions platform at genial.is/CATS-RNA-seq-demo.

  •  Applications
    DNA/RNA library preparation
    Most of the major next-generation sequencing platforms require ligation of specific adaptor oligos to fragmented DNA or RNA prior to sequencing After input DNA has been fragmented, it is end-repaired and blunt-ended. The next step is a A-tail... Read more
    Next Generation Sequencing
    DNA Shearing, library preparation, and automation: your one-stop shop for NGS 1. Choose your shearing device: Shear DNA anywhere from 150 bp to 75 kb Shear down to 5 μl: 150 bp - 2 kb Perfect for NGS DNA libr... Read more
  •  Documents
    CATS RNA-seq Kit MANUAL
    CATS RNA-seq Kit manual    		 Download
    CATS paired-end sequencing primer (for read2) DATASHEET
    This primer has been specifically designed to be used in paired-end sequencing with CATS librarie...    		 Download
    Diagenode trimming tools for CATS library preparation [read1]
    Due to special mechanisms in creating CATS libraries such as template switching and artificial po...    		 Download
    Diagenode trimming tools for CATS library preparation [read2]
    Due to special mechanisms in creating CATS libraries such as template switching and artificial po...    		 Download
    Diagenode trimming tools for CATS library preparation MANUAL
    This document describes the usage of the Diagenode trimming tools for CATS library preparation, w...    		 Download
    Material Safety Data Sheet CATS RNA-seq Kit DATASHEET
    		 Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: CATS RNA-seq Kit v2 x24 (Diagenode Cat# C05010041). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Directional high-throughput sequencing of RNAs without gene-specific primers.
    Mäki A, Tiirola M
    Ribosomal RNA analysis is a useful tool for characterization of microbial communities. However, the lack of broad-range primers has hampered the simultaneous analysis of eukaryotic and prokaryotic members by amplicon sequencing. We present a complete workflow for directional, primer-independent sequencing of size-se...

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