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  • MidRange I PFG Marker

    Description

    MidRange PFG Marker I is concatemers of λ DNA isolated from the bacteriophage λ (cI857 ind1 Sam7) mixed with XhoI digested λ DNA embedded in 1% LMP agarose and supplied in a GelSyringe™ dispenser. XhoI produces fragments of 15.0 and 33.5 kb. These fragments anneal to and form concatemers with intact λ DNA. It is designed for use as a size marker for pulsed field gel electrophoresis (PFG). Size range: 15-300 kb.



    N3551_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
    17276000
    16257500
    15242500
    14227500
    13209000
    12194000
    11179000
    10160500
    9145500
    8130500
    7112000
    697000
    582000
    463500
    348500
    233500
    115000

    Effective Size Range

    15,000bp 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 I 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. Ellard, J., Greci, J. and Davis, T.B. unpublished observation.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.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    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.
    2. Can I use SYBR® with the DNA Ladders from NEB?