Scribd Logo
Upload

Welcome to Scribd!

  • 120 views

    Quantam Freeze and Squeeze Protocol

    Uploaded by

    me_dayakar
    This document describes a method for purifying DNA fragments from agarose gels using Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Columns. The method involves excising the DNA band of interest from an agarose gel, freezing gel slices placed in a spin column filter cup, and then centrifuging to recover purified DNA in the collection tube below while retaining agarose debris in the filter. The purified DNA can then be used directly in downstream applications like PCR without further cleanup. The spin column consists of a 0.45 μm filter cup nested in a specially shaped "dolphin tube" collection tube to ensure purified DNA recovery and purity.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd

    Quantam Freeze and Squeeze Protocol

    Uploaded by

    me_dayakar
    120 views1 page
    This document describes a method for purifying DNA fragments from agarose gels using Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Columns. The method involves excising the DNA band of interest from an agarose gel, freezing gel slices placed in a spin column filter cup, and then centrifuging to recover purified DNA in the collection tube below while retaining agarose debris in the filter. The purified DNA can then be used directly in downstream applications like PCR without further cleanup. The spin column consists of a 0.45 μm filter cup nested in a specially shaped "dolphin tube" collection tube to ensure purified DNA recovery and purity.

    Copyright

    © Attribution Non-Commercial (BY-NC)

    Available Formats

    PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    This document describes a method for purifying DNA fragments from agarose gels using Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Columns. The method involves excising the DNA band of interest from an agarose gel, freezing gel slices placed in a spin column filter cup, and then centrifuging to recover purified DNA in the collection tube below while retaining agarose debris in the filter. The purified DNA can then be used directly in downstream applications like PCR without further cleanup. The spin column consists of a 0.45 μm filter cup nested in a specially shaped "dolphin tube" collection tube to ensure purified DNA recovery and purity.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    120 views1 page

    Quantam Freeze and Squeeze Protocol

    Uploaded by

    me_dayakar
    This document describes a method for purifying DNA fragments from agarose gels using Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Columns. The method involves excising the DNA band of interest from an agarose gel, freezing gel slices placed in a spin column filter cup, and then centrifuging to recover purified DNA in the collection tube below while retaining agarose debris in the filter. The purified DNA can then be used directly in downstream applications like PCR without further cleanup. The spin column consists of a 0.45 μm filter cup nested in a specially shaped "dolphin tube" collection tube to ensure purified DNA recovery and purity.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 1

    TM

    Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction


    Spin Columns
    Introduction
    Purification of double-stranded DNA fragments from TAE or TBE agarose gels with the Quantum PrepTM Freeze ‘N Squeeze DNA
    Gel Extraction Spin Column is quick and efficient. Unlike other DNA purification methods which are often time-consuming or involve
    toxic chaotropic materials, the Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Column method purifies via filtration in a
    spin column format.
    The spin column consists of a filter cup (with 0.45 µm cellose acetate filter) contained within a special 2.0 ml “dolphin” tube for
    collection of purified sample. The distinctive bottlenosed shape of the dolphin tube means recovered samples remain well below the
    bottom of the filter cup to insure purity. The extra long strap allows the cap to fit over either the filter cup during purification or to cap
    the collection tube itself for storage of the purified sample.
    In this method, a DNA band of interest is excised from an agarose gel, and the gel slice cut into small pieces and placed into the
    filter cup. The cup plus gel pieces is put in a -20 °C freezer for 5 minutes (the freeze), then removed and immediately centrifuged at
    13,000 x g for 3 minutes at room temperature (the squeeze). Agarose debris is retained within the filter cup; the liquid at the bottom of
    the tube contains the recovered DNA.
    This method allows for recovery of DNA over a wide range of fragment sizes (50 bp to 23 kbp). The recovered DNA can be used
    for PCR, ligations, labeling or other enzymatic reactions without further purification or sample preparation.

    Protocol
    1. Electrophorese the DNA sample in an agarose gel (TAE or TBE), then stain with an appropriate reagent, e.g., ethidium bromide or
    SYBRTM Green I.
    2. Using a clean razor blade, carefully excise the band of interest. Trim excess agarose from all six sides of the DNA band to maxi-
    mize recovery and purity.
    3. Chop the trimmed gel slice and place the pieces into the filter cup of the Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction
    Spin Column. Place the filter cup into the dolphin tube.
    If the volume of your trimmed gel slice is too great to fit into one filter cup, then use two or more and pool the recovered samples
    at the end of the protocol.
    4. Place the Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Column (filter cup nested within dolphin tube) in a -20° C
    freezer for 5 minutes.
    5. Spin the sample at 13,000 x g for 3 minutes at room temperature.
    6. Collect the purified DNA from the collection tube; the agarose debris will be retained within the filter cup of the Quantum Prep
    Freeze ‘N Squeeze DNA Gel Extraction Spin Column. The DNA is ready to use for PCR, ligations, labeling or other enzymatic
    reactions. Ethanol precipitation is recommended for applications requiring a more concentrated sample and will also have the effect
    of further purifying the sample.

    References
    1. Thuring, R.W.J., Sanders, J.P.M. and Borst, P., Anal. Biochem., 66, 213, (1975).

    Product Information
    Catalog
    Number Description
    732-6165 Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Column, 25 per bag
    732-6166 Quantum Prep Freeze ‘N Squeeze DNA Gel Extraction Spin Column, 100 per bag
    732-6160 Quantum Prep Gel Slice Kit
    4106139 Rev B

    You might also like