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    Cat. No.
    Size
    List Price*
    QTY
    BIO-52068
    10 preps
    $42.50
    +
    BIO-52069
    50 preps
    $176.00
    +
    BIO-52070
    250 preps
    $784.00
    +
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    Description

    Highly efficient purification of genomic DNA by combining the stringency of either CTAB based lysis for plants rich in polyphenolics, polysaccharides or SDS based lysis for plants rich in polyphenolics or polysaccharides, with the speed and purity of silica-membrane column purification.

    Product Highlights

    • Fast – streamlined protocol for reliable recovery of genomic DNA from six samples in as little as 30 minutes
    • High-performance – extraction of high-quality DNA, ideal for use in all downstream applications
    • Efficient – optimized lysis conditions and column matrix for improved recovery of genomic DNA from a wide range of plant samples
    • Versatile – a choice of two lysis buffers ensures excellent genomic DNA recovery from any plant material
    • Convenient – kit includes all necessary components, including filters (shredders) and RNase A
    • Safe - no hazardous phenol/chloroform extraction, CsCl centrifugation, LiCl or alcohol precipitation

    Product Description

    The ISOLATE II Plant DNA Kit provides a simple, efficient column-based method for the isolation of genomic DNA from a wide variety of plant materials, without the need for hazardous reagents such as phenol.

    By combining the stringency of CTAB lysis with the speed and ease-of-use of silica-membrane purification, the ISOLATE II Plant DNA Kit provides a fast method for the purification of high-quality genomic DNA from most plant cells, particularly those rich in polysaccharides, including leaves, bark, roots and fruits as well as dung, animal-fecal, soil and compost samples.

    Some plant tissues do not lyse well in CTAB and so an SDS-based lysis buffer is also provided as an alternative. Residual amounts of RNA remaining can be removed using an RNase treatment during lysis with RNase A that is supplied with the kit. 

    The ISOLATE II Plant DNA Kit has been designed to deliver optimal performance in qPCR in tandem with the SensiFAST Real-Time PCR Kits and in end-point PCR with any enzyme from the Bioline PCR portfolio, including MyTaq DNA Polymerase

    Applications

    • qPCR
    • End-point PCR
    • Southern, dot and slot blotting
    • Genotyping
    • Restriction digestion
    • Next Generation Sequencing
    • Bisulfite conversion/methylation analysis
    Main
    Fast purification of high-quality genomic DNA from a variety of plant tissues, including leaves, bark, roots and fruits.

    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

    Application Note

    ISOLATE II Plant DNA Kit

    Isolating DNA from soil samples is a big challenge because samples are full of inhibitors like polymers or low-molecular substances, which inhibit downstream applications. Therefore, it is difficult to find a kit which can deliver stable DNA. The ISOLATE II Plant DNA Kit is able to handle these kinds of samples and worked well for soil samples.

    Katharina Schulte, Technical College Zweibruecken, Germany

    Product Selection

    Please refer to the ISOLATE II Selection Chart to confirm the applications for which the ISOLATE II Plant DNA Kit is recommended.


    Specification

    Components

    Reagent

    10 Preps

    50 Preps

    250 Preps

    ISOLATE II Filters

    10

    50

     250

    ISOLATE II Plant Columns

    10

    50

     250

    Collection Tubes (2 mL)

    20

    100

    500 

    Lysis Buffer PA1

    5 mL

    25 mL

    125 mL

    Lysis Buffer PA2

    4 mL

    20 mL

    100 mL

    Precipitation Buffer PL3

    1 mL

    10 mL

    25 mL

    Binding Buffer PB

    6 mL

    30 mL

    125 mL

    Wash Buffer PAW1

    6 mL

    30 mL

    125 mL

    Wash Buffer PAW2

    6 mL

    25 mL

    50 mL

    Elution Buffer PG

    13 mL

    13 mL

    30 mL

    RNase A (lyophilized)

    1.5 mg

    6 mg

    2 x 15 mg

    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

    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.

    Both lysis buffers PA1 and PA2 can possibly become cloudy, if they are stored under 20°C. They contain CTAB (PA1) and SDS (PA2) which can precipitate. After a short incubation at 37°C for several minutes they should become clear again.



    The best lysis buffer for distinct samples is not predictable, try both lysis buffer (SDS/CTAB) side by side to determine the optimal buffer for your plant sample.

    The ISOLATE II Genomic DNA Kit provides a protocol for hard to lyse bacteria (see sec 9.2). Also the ISOLATE II Plant DNA kit could be suitable, because the optimized lysis buffer from this kits may be also be used for processing these difficult to lyse bacteria. In general, it is necessary to optimize the lysis step with the appropriate mechanical, enzymatic or chemical methods recommended for this kind of sample. Please refer to the related literature or ask the Bioline technical support for further suggestion.

    It is possible to use the ISOLATE II Plant DNA and RNA kits. The optimized lysis of these kits may be also useful for processing difficult to lyse bacteria and fungi in soil samples

    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.



    Reviews

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