Diagenode

CATS RNA-seq Kit v2 x24

カタログ番号
フォーマット
価格
C05010041
24 rxns
$1,410.00
ブローカー情報

DiagenodeのCATS RNA-seq Kitは、RNAの低入力量から次世代シーケンシングに使用するDNAライブラリーを作製するためのライゲーションフリーの方法である革新的な "捕捉と増幅による尾引きと切り換え"(CATS)を利用しています。

  • より高いRNAの多様性
  • 10pg以下の超低入力で優れた再現性
  • 複雑なサンプルでの優れたパフォーマンス
  • 続きを読む

    CATSで核酸3末端を補足する際のポリヌクレオチドテーリング、またはcDNA合成する5'末端を捕捉する際のMMLV-RTのテンプレートスイッチング能力が非常に重要になります。このキットなら、わずか数時間で10ピコグラム以下のRNA入力からIllumina用互換のシーケンシング準備済ライブラリーを作成することができます。.

    ライブラリーは、起源の鎖特異性を保持します。他社のソリューションとは異なり、当社のCATS RNA-seqキットはライゲーションベースの方法と比較してもより高いライブラリー効率を示し、より良いテンプレート捕捉からさらに高い複雑性、より少ない増幅条件による最小バイアスをご提供します。

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

  • 特徴
    • 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.

  •  資料
    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
  •  出版物

    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...

  •  関連商品

イベント

  • Japanese Biochemical Society
    Yokohama
    Sep 18-Sep 20, 2019
  • The 2019 PacBio Long-Read Revolution: Highly Accurate and Affordable SMRT Sequencing
    Atlanta, Georgia
    Sep 19, 2019
  • The 2019 PacBio Long-Read Revolution: Highly Accurate and Affordable SMRT Sequencing
    RTP, North Carolina
    Sep 20, 2019
  • EpiGeneSys
    London
    Sep 22-Sep 24, 2019
 すべてのイベントを見る

         Site map   |   Contact us   |   Conditions of sales   |   Conditions of purchase   |   Privacy policy   |   Diagenode Diagnostics