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    Cat. No.
    Size
    List Price*
    QTY
    BIO-52087
    50 Preps
    $585.00
    +
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    Description

    Highly efficient purification of total RNA and genomic DNA by combining the stringency of guanidinium thiocyanate lysis with the speed and purity of silica-membrane column purification.

    Product Highlights

    • Reliable – excellent recovery RNA and DNA from a single FFPE sample with no splitting of the lysate, thereby reducing variability and improving consistency in results
    • Efficient – effective reversal of formalin crosslinks for improved nucleic acid yield and purity
    • Versatile – optimized for recovery of all DNA above 80 base pairs in length and all RNA types, from large mRNA species to small miRNA molecules
    • Safe - no hazardous phenol/chloroform extraction, CsCl centrifugation, LiCl or alcohol precipitation

    Product Description

    The ISOLATE II FFPE RNA/DNA Kit provides a simple, efficient column-based method for the isolation of total RNA and genomic DNA from a wide variety of starting materials, without the need for hazardous reagents such as phenol.

    By combining the stringency of guanidinium-thiocyanate lysis with the speed and ease-of-use of silica-membrane purification, the ISOLATE II FFPE RNA/DNA Kit provides a fast method for the purification of high-quality total RNA and genomic DNA from formalin-fixed paraffin-embedded (FFPE) tissue samples.

    Using formalin to fix tissues leads to crosslinking of the nucleic acids and proteins and the process of embedding the tissue samples can also lead to fragmentation of the nucleic acids over time. The ISOLATE II FFPE RNA/DNA Kit provides conditions that allow for the partial reversal of the formalin modifications, resulting in a high quality and yield of nucleic acids.

    The ISOLATE II FFPE RNA/DNA Kit has been designed to deliver optimal performance in qPCR in conjunction with either the SensiFAST cDNA Synthesis Kit and SensiFAST Real-Time PCR Kits, or the SensiFAST One-Step Real-Time RT-PCR Kits. Additionally, the ISOLATE II FFPE RNA/DNA Kit can be used to purify samples prior to PCR and RT-PCR amplification using the Tetro cDNA Synthesis Kit and any enzyme from the Bioline PCR portfolio, including MyTaq DNA Polymerase.

    Applications

    • qPCR and RT-qPCR
    • PCR and RT-PCR
    • Next Generation Sequencing
    • Genotyping
    • Northern and Southern, dot and slot blotting
    • Array analysis
    • Restriction digestion
    • Bisulfite conversion/methylation analysis
    Main
    Fast purification of high-quality total RNA and genomic DNA from formalin fixed paraffin embedded samples

    Nucleic Acid Isolation Guide

    Download the ISOLATE II Guide with detailed product descriptions and performance data to help you choose the best product for your research

    Resources

    Certification of Analysis (COAs)

    Specification

    Components

    Reagent 50 Preps
    ISOLATE II FFPE RNA Micro Columns 50
    ISOLATE II FFPE DNA Micro Columns 50
    Collection Tubes (2 mL) 100
    Elution Tubes (1.7 mL) 100
    Proteinase K (lyophilized) 2 x 12 mg
    Digestion Buffer DX 2 x 25 mL
    Lysis Buffer RX 40 mL
    Wash Buffer W1 2 x 38 mL
    RNA Elution Buffer 6 mL
    DNA Elution Buffer 15 mL
    DNase I Solution (RNase-free) 210 µL
    DNase I Reaction Buffer DRB 6 mL
    Product Manual 1
    Bench Protocol Sheet 1

    Storage & Stability

    The DNase I Solution and Proteinase K should be stored at -20°C upon arrival. All other components should be stored dry and at room temperature.

    When stored under the recommended conditions and handled correctly, full activity of reagents is retained until the expiry date indicated on the outer box label.

    Shipping conditions

    Ambient temperature



    FAQs

    The columns supplied with the ISOLATE II kits may appear similar, but each type has been optimized to work within the buffer system supplied with the corresponding kit. The swapping of columns (or buffers) between kits may lead to no recovery of nucleic acid whatsoever, or at the very least severely impaired purification.

    Interruption of the DNA/RNA extraction process is possible after the sample lysis only. It is possible to homogenize and lyse the samples and to store them in the freezer until use for RNA extraction. RNA clean up with a column cannot be interrupted and we recommend to avoid delays during the column purification process. If a delay is unavoidable the columns should be stored on ice.

    The elution buffer is ideal for applications such as restriction enzyme digestion, sequencing and PCR. It is possible to use DNase-free water for elution, but you should expect a slightly lower yield.

    For the optimal recovery of large RNA/DNA fragments it is necessary to optimize the elution step. To increase the recovery rate it would be helpful to use a larger volume for elution, incubate the column with the elution buffer prior to centrifugation and use repeated elution steps. To avoid excessive dilution it may be helpful to reapply the eluted fraction again. Furthermore it could be helpful for DNA elution to heat the elution buffer to 70°C.