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    Cat. No.
    Size
    List Price*
    QTY
    BIO-52058
    10 Preps
    £30.00
    +
    BIO-52059
    50 preps
    £88.00
    +
    BIO-52060
    250 preps
    £373.00
    +
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    Description

    Simple purification of DNA fragments, including PCR products, from agarose gels with the speed and purity of silica-membrane purification.

    Product Highlights

    • Fast - streamlined protocol for simple recovery of DNA from up to six gel slices in as little as 10 minutes
    • Efficient - column matrix for recovery rates of between 70% and 95%
    • High-performance – extraction of consistently high quality DNA fragments, ideal for use in all downstream applications
    • Versatile - single buffer promotes effective DNA recovery from both TAE and TBE agarose gels
    • Safe - no hazardous phenol/chloroform extraction, CsCl centrifugation, LiCl or alcohol precipitation

    Product Description

    The ISOLATE II PCR and Gel Kit provides a simple, efficient membrane-based method for the purification of DNA from PCR reactions and from TAE and TBE agarose gels, without the need for hazardous reagents or ethanol precipitation.

    Six PCR products can be purified in 10 minutes using simple binding and elution steps. Concentrated PCR products ranging between 60 bp and 15 kb can be eluted, removing primers, nucleotides, enzymes, mineral oil, salts and other impurities.

    DNA fragments between 50 bp and 20 kb can be extracted from six agarose gel slices in about 20 minutes using a color indicator to help maintain optimal pH and identify undissolved agarose.

    The ISOLATE II PCR and Gel Kit has been designed to deliver optimal performance in transformations, cloning, sequencing and restriction analysis.

    Applications

    • Cloning
    • Ligation
    • Restriction digestion
    • Fluorescence sequencing
    • Labeling
    • PCR
    • Transfection
    • In vitro transcription
    Main
    Fast purification of DNA from PCR reactions and agarose gels.

    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

    Specification

    Components

    Reagent

    10 Preps

    50 Preps

    250 Preps

    ISOLATE II PCR and Gel Columns (yellow)

    10

    50

    250

    Collection Tubes

    10

    50

    250

    Binding Buffer CB

    10 mL

    40 mL

    200 mL

    Wash Buffer CW (Concentrate)

    6 mL

    25 mL

    2 x 50 mL

    Elution Buffer C

    13 mL

    13 mL

    30 mL

    Bench Protocol Sheet

    1

    1

    1

    Storage & Stability

    All 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

    Shipped at 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.

    The elution buffer of this kit does not contain EDTA which could interfere with downstream applications like PCR or sequencing.

    Yes, though this may require some optimization. Warming the elution buffer to 50°C or even 70°C before applying it to the column can help to elute long PCR products, but there is the possibility using this adaptation to the protocol that the primers will be eluted too.



    The ISOLATE II PCR and Gel Kit can be used under standard conditions to separate most PCR primers away from their product. A primer of only 30 bp is efficiently separated, but longer mutagenic primers of around 100 bp may be separated from the PCR buffer along with the amplicon.

    The ISOLATE II Plasmid Mini Kit can be used to both purify and concentrate naked plasmid DNA. The DNA can be loaded directly onto a fresh column then the remaining washing, drying and elution steps followed as for the standard protocol. The sample can be concentrated by reducing the elution volume in the final stage. However, a better recovery can be obtained by using the ISOLATE II PCR and Gel Kit and treating the plasmid sample as though it were a PCR product.

    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.