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  • MidRange II PFG Marker

    Discontinued Date

    01/01/2013
    Catalog #SizeConcentrationGel LanesPriceQtyAdd to Cart
      
    Categories:
    Discontinued Products

    Description

    MidRange Marker II is concatemers of λ DNA isolated from the bacteriophage λ (cI857 ind1 Sam7) mixed with XbaI digested λ DNA embedded in 1% LMP agarose and supplied in a GelSyringe™ dispenser. XbaI produces fragments of 24.0 and 24.5 kb. These fragments anneal to and form concatamers with intact λ DNA. It is designed for use as a size marker for pulsed field gel electrophoresis (PFG). Size range: 24-300 kb.



    N3552_thumb

    1% agarose gel, 6 V/cm, 15°C for 24 hours. Switch times ramped from 1-25 seconds.

    Properties and Usage

    Bases

    Fragmentbp
    18291000
    17267000
    16266500
    15242500
    14218500
    13218000
    12194000
    11170000
    10169500
    9145500
    8121500
    7121000
    697000
    573000
    472500
    348500
    224500
    124000

    Effective Size Range

    24,000bp to 291,000bp

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    1 mM EDTA
    50% Glycerol
    1% LMP agarose
    pH 8.0 @ 25°C

    Notes

    1. The photograph represents the pulsed field gel separation of Mid Range II Markers using a CHEF apparatus. The 1% agarose gel was run at 6 volts/cm using ramped pulse times from 1 to 25 seconds for 24 hours at 15°C in 0.5X TBE (50 mM Tris-HCl, 50 mM boric acid, 1 mM EDTA) made with Milli-Q™ water.
    2. Extrude agarose from GelSyringe carefully and slice plugs from the end with a sharp blade. One plug is sufficient for one lane of a gel. Place the plug at the front of the well and seal with molten agarose. Allow no bubbles to form.
    3. Recommended plug sizes are from 5-10 µl. A 10 µl plug (one small graduation on the GelSyringe volume scale) contains approximately 0.5 µg of DNA. Each GelSyringe yields 50+ plugs.
    4. Melting plugs will cause denaturation of concatemers.
    5. Place plug at the front of the well and seal with molten agarose just above gelling temperature(~42–45°C). Allow no bubbles to form. Never attach the agarose plugs to the gel comb before the gel is poured. Heat from the solidifying gel will cause the Lambda concatemers to denature.

    References

    1. Greci, J. and Davis, T.B., New England Biolabs, unpublished observation.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.
    1. There are several factors that lead to poor results for PFG markers. Here is a list of recommendations for ensuring a sharp high quality banding pattern.