Diagenode

Tagmentase (Tn5 transposase) - loaded

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目录号
格式
C01070012-200
200 μl
其他格式

Diagenode Tagmentase – loaded is a hyperactive Tn5 transposase preloaded with Illumina-compatible sequencing adapters. Its ability to cut DNA and insert the sequencing adapters in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq or ChIPmentation.

Using Diagenode’s Tagmentase (Tn5 transposase) - loaded you may need also:

Looking for unloaded Tagmentase? Please go to Tagmentase (Tn5 transposase) – unloaded

TESTIMONIAL

We have been using the Hyperactive Tagmentase for 2 years and its performance is outstanding - short operation time and good reproducibility, outmatching the competition. Moreover the interaction with customer representatives is always top-notch - highly efficient and knowledgeable. I can't recommend enough!

Julia Liz Touza, AstraZeneca Gothenburg, Sweden
  • Examples of results

    Tagmentase (Tn5 transposase) – loaded can be used in any application using tagmentation-based library preparation, like for example ChIPmentation, ATAC-seq.  Here below we are showing some examples of results.

    Tn5 transposase Enxzymes

    Figure 1. Typical library profile of ATAC-seq library generated with the Tagmentase
    ATAC-seq has been performed on 50,000 nuclei from K562 cells using the Tagmentase (Tn5 transposase) – loaded (Cat. No. C01070012) and Tagmentation Buffer (2x) (Cat. No. C01019043). The library profile has been generated on a Fragment Analyzer (Agilent).

    Diagenode Tagmentation

    Figure 2. Enrichments at TSS of ATAC-seq libraries generated with the Tagmentase
    ATAC-seq has been performed on 50,000 nuclei from K562 cells, in triplicates, using the Tagmentase (Tn5 transposase) – loaded (Cat. No. C01070012) and Tagmentation Buffer (2x) (Cat. No. C01019043). Heatmaps around the hg19 TSS were generated using deeptools plotHeatmap functionality.

    Transposase enzymes for ATAC-Seq

    Figure 3. Sequencing profiles of ATAC-seq libraries generated with the Tagmentase
    ATAC-seq has been performed on 50,000 nuclei from K562 cells, in triplicates, using the Tagmentase (Tn5 transposase) – loaded (Cat. No. C01070012) and Tagmentation Buffer (2x) (Cat. No. C01019043).

    purified Transposase enzymes

    Figure 4. Typical library profile of ChIPmentation library generated with the Tagmentase
    Chromatin preparation and immunoprecipitation have been performed on 1,000,000 K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011010) in combination with the Diagenode antibody targeting H3K4me3 (Cat. No. C15410003) and with the Tagmentase (Tn5 transposase) – loaded (Cat. No. C01070012). The library profile has been generated on a Fragment Analyzer (Agilent).

    pA-Tn5 Transposase Enzymes

    Figure 5. Sequencing profiles of ChIPmentation libraries generated with the Tagmentase
    Chromatin preparation and immunoprecipitation have been performed on 500.000 to 1,000,000 cells using the ChIPmentation Kit for Histones (Cat. No. C01011010) in combination with the Tagmentase (Tn5 transposase) – loaded (Cat. No. C01070012) on K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K36me3 (Cat. No. C15410192), H3K27me3 (Cat. No. C15410195) and rabbit IgG (Cat. No. C15410206) have been used.

  • Product information

    Tagmentase (Tn5 transposase) - loaded 

    Protein Molecular weight: 53.3 kDa

    Expressed: in Escherichia coli

    Product description:  Diagenode Tagmentase – loaded is a hyperactive Tn5 transposase preloaded with Illumina-compatible sequencing adapters. Its ability to cut DNA and insert the sequencing adapters in one step makes it the perfect companion for Next-Generation Sequencing experiments. The Tagmentase is pre-loaded with sequencing adapters compatible with Illumina Nextera platforms, as shown below. The oligos loaded on the Tagmentase are inserted into DNA upon a tagmentation reaction.

    Mosaic end_reverse: 5’ [PHO]CTGTCTCTTATACACATCT 3’
    Mosaic end_Adapter A: 5’ TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG 3’

    Mosaic end_Adapter B: 5’ GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG 3’

    Underlined regions correspond to the double-stranded part of the adapter, recognized by the Tagmentase. The final libraries can be amplified using Diagenode Primer indexes for tagmented libraries:
    24 SI for tagmented libraries, Cat. No. C01011032
    24 UDI fortagmented libraries - Set I, Cat. No. C0101134
    24 UDI for tagmented libraries - Set II, Cat. No. C0101136

    24 UDI for tagmented libraries - Set III, Cat. No. C0101137

    Storage conditions: Store at -20°C. Guaranteed stable for 6 months from date of receipt when stored properly.

    Storage buffer: Supplied in solution containing 50% v/v glycerol.

    Properties & Usage: Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.

    Applications: Tagmentase (Tn5 transposase) – loaded can be used in a variety of applications to construct library for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.

    For ATAC-seq and ChIPmentation, we recommend using Diagenode validated protocols:
    ATAC-seq kit, Cat. No. C01080002
    ATAC-seq package for tissue, Cat. No. C01080006
    ChIPmentation Kit for Histones, Cat. No. C01011009
    µChIPmentation Kit for Histones, Cat. No. C01011011
    TAG Kit for ChIPmentation, Cat. No. C01011030
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    How to properly cite this product in your work

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    Using our products in your publication? Let us know!

    ARMC5 selectively degrades SCAP-free SREBF1 and is essential for fatty acid desaturation in adipocytes
    Akifumi Uota et al.
    SREBF1 plays the central role in lipid metabolism. It has been known that full-length SREBF1 that did not associate with SCAP (SCAP-free SREBF1) is actively degraded, but its molecular mechanism and its biological meaning remain unclear. ARMC5-CUL3 complex was recently identified as E3 ubiquitin ligase of full-lengt...

    HNF1β bookmarking involves Topoisomerase 1 activation and DNA topology relaxation in mitotic chromatin
    Alessia Bagattin et al.
    Highlights HNF1β mitotic site binding is preserved with a specific methanol/formaldehyde ChIP BTBD2, an HNF1β partner, mediates mitosis-specific interaction with TOP1 HNF1β recruits TOP1 and induces DNA relaxation around bookmarked HNF1β sites An HNF1β m...

    Nuclear lamin A/C phosphorylation by loss of androgen receptor leads to cancer-associated fibroblast activation
    Ghosh S. et al.
    Alterations in nuclear structure and function are hallmarks of cancer cells. Little is known about these changes in Cancer-Associated Fibroblasts (CAFs), crucial components of the tumor microenvironment. Loss of the androgen receptor (AR) in human dermal fibroblasts (HDFs), which triggers early steps of CAF act...

    A critical role for HNF4α in polymicrobial sepsis-associated metabolic reprogramming and death
    van Dender C. et al.
    In sepsis, limited food intake and increased energy expenditure induce a starvation response, which is compromised by a quick decline in the expression of hepatic PPARα, a transcription factor essential in intracellular catabolism of free fatty acids. The mechanism upstream of this PPARα downregulation i...

    High-throughput sequencing of insect specimens with sub-optimal DNA preservation using a practical, plate-based Illumina-compatible Tn5 transposase library preparation method
    Cobb L. et all.
    Entomological sampling and storage conditions often prioritise efficiency, practicality and conservation of morphological characteristics, and may therefore be suboptimal for DNA preservation. This practice can impact downstream molecular applications, such as the generation of high-throughput genomic libraries, whi...

    On the identification of differentially-active transcription factors from ATAC-seq data
    Gerbaldo F. et al.
    ATAC-seq has emerged as a rich epigenome profiling technique, and is commonly used to identify Transcription Factors (TFs) underlying given phenomena. A number of methods can be used to identify differentially-active TFs through the accessibility of their DNA-binding motif, however little is known on the best approa...

    Cellular reprogramming in vivo initiated by SOX4 pioneer factor activity
    Katsuda T.
    Tissue damage elicits cell fate switching through a process called metaplasia, but how the starting cell fate is silenced and the new cell fate is activated has not been investigated in animals. In cell culture, pioneer transcription factors mediate “reprogramming” by opening new chromatin sites for expr...

    The ncBAF complex regulates transcription in AML through H3K27ac sensing by BRD9
    Klein D.C. et al.
    The non-canonical BAF complex (ncBAF) subunit BRD9 is essential for acute myeloid leukemia (AML) cell viability but has an unclear role in leukemogenesis. Because BRD9 is required for ncBAF complex assembly through its DUF3512 domain, precise bromodomain inhibition is necessary to parse the role of BRD9 as a transcr...

    ARID1A governs the silencing of sex-linked transcription during male meiosis in the mouse
    Menon D.U. et al.
    We present evidence implicating the BAF (BRG1/BRM Associated Factor) chromatin remodeler in meiotic sex chromosome inactivation (MSCI). By immunofluorescence (IF), the putative BAF DNA binding subunit, ARID1A (AT-rich Interaction Domain 1a), appeared enriched on the male sex chromosomes during diplonema of meiosis I...

    Zfp296 knockout enhances chromatin accessibility and induces a uniquestate of pluripotency in embryonic stem cells.
    Miyazaki S. et al.
    The Zfp296 gene encodes a zinc finger-type protein. Its expression is high in mouse embryonic stem cells (ESCs) but rapidly decreases following differentiation. Zfp296-knockout (KO) ESCs grew as flat colonies, which were reverted to rounded colonies by exogenous expression of Zfp296. KO ESCs could not form teratomas...

    YAP/BRD4-controlled ROR1 promotes tumor-initiating cells andhyperproliferation in pancreatic cancer.
    Yamazaki M. et al.
    Tumor-initiating cells are major drivers of chemoresistance and attractive targets for cancer therapy, however, their identity in human pancreatic ductal adenocarcinoma (PDAC) and the key molecules underlying their traits remain poorly understood. Here, we show that a cellular subpopulation with partial epithelial-m...

    Analyzing genomic and epigenetic profiles in single cells by hybridtransposase (scGET-seq).
    Cittaro D. et al.
    scGET-seq simultaneously profiles euchromatin and heterochromatin. scGET-seq exploits the concurrent action of transposase Tn5 and its hybrid form TnH, which targets H3K9me3 domains. Here we present a step-by-step protocol to profile single cells by scGET-seq using a 10× Chromium Controller. We describ...

    A neurodevelopmental epigenetic programme mediated bySMARCD3-DAB1-Reelin signalling is hijacked to promote medulloblastomametastasis.
    Zou Han et al.
    How abnormal neurodevelopment relates to the tumour aggressiveness of medulloblastoma (MB), the most common type of embryonal tumour, remains elusive. Here we uncover a neurodevelopmental epigenomic programme that is hijacked to induce MB metastatic dissemination. Unsupervised analyses of integrated publicly availab...

    Physiological reprogramming in vivo mediated by Sox4 pioneer factoractivity
    Katsuda T. et al.
    Tissue damage elicits cell fate switching through a process called metaplasia, but how the starting cell fate is silenced and the new cell fate is activated has not been investigated in animals. In cell culture, pioneer transcription factors mediate “reprogramming” by opening new chromatin sites for expr...

    EBF1 is continuously required for stabilizing local chromatinaccessibility in pro-B cells.
    Zolotarev Nikolay et al.
    The establishment of de novo chromatin accessibility in lymphoid progenitors requires the "pioneering" function of transcription factor (TF) early B cell factor 1 (EBF1), which binds to naïve chromatin and induces accessibility by recruiting the BRG1 chromatin remodeler subunit. However, it remains unclear whet...

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