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产品英文名:bisulflash dna modification kit
产品中文名:
产品编号:P-1026-050      产品品牌:epigentek备有现货
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产品详细

Product Overview
  • Product Details
  • Product Components
  • User Guide & MSDS
  • Product Citations
  • Product Ovrview

    The BisulFlash™ DNA Modification Kit is a complete set of optimized buffers and reagents to perform DNA modification using a next generation DNA bisulfite conversion technology developed by Epigentek. Through a proprietary composition which allows DNA denaturation and bisulfite conversion to be processed at the same time, the complete procedure is reduced to only 30 minutes. Furthermore, it prevents more than 90% of DNA loss, completely converting unmethylated cytosine into uracil.

    Perfecting Bisulfite Conversion
    Traditional methods involve a separate denaturation step followed by a subsequent sodium bisulfite DNA conversion step -- but with the BisulFlash™ method, DNA denaturation status is concurrently sustained throughout the entire bisulfite DNA conversion process. This breakthrough approach enables the DNA conversion process to be sigificantly faster with higher conversion efficiency and accuracy. We continue to innovate with the development of the new BisulFlash™ kit by identifying four critical components of bisulfite conversion:
    • Speed: Reduce the entire procedure to as short as 30 minutes without any reagent setup time.
    • Efficiency: Completely convert unmethylated cytosine into uracil -- modified DNA > 99.99%.
    • DNA Protection: Protect against DNA degradation of which more than 90% of DNA loss can be prevented, allowing for greater recovery.
    • Sensitivity: Start with the lowest amount of input DNA for modification -- only 0.2 ng or just 50 cells.
    The convenient ready-to-use, DNA conversion mix solution and single temperature incubation along with the features mentioned above allow for true perfection in bisulfite conversion. The BisulFlash™ DNA Modification Kit is suitable for bisulfite sequencing, MS-PCR, real time MS-PCR, and methylation microarray.
    Comparative Overview of Commercial Kits
    Data was obtained through actual use of the kit, customer feedback, or information provided by the supplier's datasheet or website.

     BisulFlashSupplier 1Supplier 2Supplier 3
    Processing Time30 min>3 hours>6 hours12-16 hours
    Correct Conversion99.99%99.5%99.4%98%
    Error Conversion<0.1%>0.3%N/A>1.5%
    Correct/Error Conversion Ratio≅ 1000≅ 330N/A≅ 65
    DNA DegradationVery lowMediumLowHigh
    Minimum Starting DNA0.2 ng0.5 ng1 ng1 ng
    ConvenienceVery highHighMediumLow

    Fig. 1. Schematic procedure of the BisulFlash™ DNA Modification Kit to obtain converted DNA.
    Product Details

    Principle & Procedure
    As a next generation bisulfite conversion tool, the BisulFlash™ DNA Modification Kit contains all reagents required for an ultra-fast bisulfite conversion on a DNA sample. With the ready-to-use convesion mix solution, DNA denaturation status is sustained throughout the entire bisulfite DNA conversion process. This proprietary solution allows the bisulfite reagents to rapidly convert all cytosine to uracil with negligible methylcytosine conversion. The unique DNA protection reagents contained in the mix can prevent the chemical and thermophilic degradation of DNA in the bisulfite treatment. The non-toxic DNA capture solution enables DNA to tightly bind to the column filter, thus DNA cleaning can be carried out on the column to effectively remove residual bisulfite and salts.

    Rapid Results
    Only 30 minutes are required for the entire BisulFlash™ procedure -- from converting your sample DNA to pure bisulfite modified DNA. That is significantly faster than any currently used kits (2-8 h) or homebrew methods (>16 h). The BisulFlash™ kit provides everything required for a successful bisulfite conversion and DNA cleanup in the shortest time with the fewest steps posible.

    Perfect DNA Conversion
    Each reaction with the BisulFlash™ kit can use 0.2 ng – 1 µg of DNA. For optimal conversion, the DNA amount is 200-500 ng. The novel procedure and proprietary ready-to-use DNA conversion mix solution allow DNA denaturation and bisulfite conversion to occur at the same time and enable all cytosines to be converted to uracil (>99.999%), while 5-methylcytosine remains the same. The highly efficient cytosine conversion is proven by the low CT values obtained when amplifying converted DNA using real-time PCR (Fig. 2). This perfect conversion rate is superior to other bisulfite kits available on the market and provides repeatable and dependable downstream analysis.

    Powerful DNA Protection
    DNA protection reagents are added into the DNA conversion mix solution to prevent DNA from chemical and thermophilic degradation in the bisulfite treatment and provide effective DNA denaturation, resulting in single-tranded DNA necessary for complete cytosine conversion. The prevention of DNA degradation enables subsequent amplification and analysis of large PCR fragments (Fig. 3). The efficient integrated DNA cleanup and unique elution buffer allow for long-term storage (> 6 months) and multiple freezing/thawing of the converted DNA without affecting the DNA quality.


    Fig. 2. Complete Cytosine Conversion: 200 ng of genomic DNA isolated from 3 cancer cell lines was treated with the BisulFlash™ DNA Modification Kit. Next, the unconverted and converted DNA in each treated sample were determined using unconverted DNA-specific and converted DNA-specific primers (β-actin, 110 bps), respectively. A: real time PCR; B: end-point PCR. The BisulFlash&trade kit treated DNA was completely converted, and no unconverted DNA in the treated samples was determined after 45 cycles.

    Fig. 3. Effective DNA Protection: Fully methylated human genomic DNA at various amounts (0.2 ng-200 ng) were converted using the BisulFlash™ DNA Modification Kit. 1 µl of 20 µl eluate was used for real time qPCR and a pair of primers was used to amplify converted DNA. As little as 0.2 ng DNA is sufficient for bisulfite conversion using the BisulFlash™ DNA Modification Kit. A: real time PCR; B: starting DNA amount-CT value curve.
    Product Components

    BF1 (Conversion Mix Solution)br />BF2 (Capture Solution)
    BF3 (Desulphonation Solution)
    BF4 (Elution Solution)
    BF5 (Conversion Enhancer)
    BF6 (Denaturation Enhancer)
    F-Spin Columns
    F-Collection Tubes
    User Guide

    User Guide & MSDS

    [User Guide]*
    *Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by ing along with your contact information and institution name.

    Product Citations

    Mrtínez-Macías, MI. et. al. (February 2012). A DNA 3' phosphatase functions in active DNA demethylation in Arabidopsis. Mol Cell. 45 (3):357-70. PubMed Abstract

    Yang, J. et. al. (November 2011). Stem Cell Gene SALL4 Suppresses Transcription through Recruitment of DNA Methyltransferases.J Biol Chem. Epub ahead of print. PubMed Abstract

    Yang, X. et. al. (September 2011). Neurodegenerative and Inflammatory Pathway Components linked to TNF-alpha-TNFR1 Signaling in the Glaucomatous Human Retina. Invest Ophthalmol Vis Sci. Epub ahead of print. PubMed Abstract

    Geyer, K. et. al. (August 2011). Cytosine methylation regulates oviposition in the pathogenic bood fluke Schistosoma mansoni. Nat Commun. 2:424. PubMed Abstract

    Komori, HK. et. al. (August 2011). Application of microdroplet PCR for large-scale targeted bisulfite sequencing. Genome Res. Epub ahead of print. PubMed Abstract

    Carrard, A. et. al. (August 2011). Increased DNA methylation status of the serotonin receptor 5HTR1A gene promoter in schizophrenia and bipolar disorder. J Affect Disord. 132(3):450-3. PubMed Abstract

    Cho, YH. et. al. (July 2011). Identification of transcriptional regulatory elements required for the Mup2 expression in circadian clock mutant mice. Biochem Biophys Res Commun. 410(4):834-40. PubMed Abstract

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