Diagenode

CATS RNA-seq Kit v2 x12

Catalog Number
Format
Price
C05010045
12 rxns
$780.00
Other format

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.

  •  Documents
    CATS RNA-seq Kit MANUAL
    CATS RNA-seq Kit manual
    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 x12 (Diagenode Cat# C05010045). Click here to copy to clipboard.

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    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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