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  • Lambda Ladder PFG Marker

    Description

    Successively larger concatemers of lambda DNA (cI857 ind 1 Sam7) embedded in 1% LMP agarose and supplied in a GelSyringe™ dispenser. Designed to be used as size markers for pulsed-field gel electrophoresis (PFG). Size range: 50-1,000 kb.

    N0340_thumb

    1% agarose gel, 4.5 V/cm, 15°C for 48 hours. Switch times ramped from 5-120 seconds.

    Properties and Usage

    Bases

    Fragmentbp
    211018500
    20970000
    19921500
    18873000
    17824500
    16776000
    15727500
    14679000
    13630500
    12582000
    11533500
    10485000
    9436500
    8388000
    7339500
    6291000
    5242500
    4194000
    3145500
    297000
    148500

    Effective Size Range

    48,500bp to 1,018,500bp

    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 Lambda Ladders using a CHEF apparatus. The 1% agarose gel was run at 4.5 volts/cm using ramped pulse times from 5 to 120 seconds for 48 hours at 15°C in 0.5X TBE (50 mM Tris-HCl, 50 mM boric acid, 1 mM EDTA) made with Milli-Q™ water allowing resolution of 20 Lambda Ladder bands.
    2. 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 25+ plugs.
    3. 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.
    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. Burke-Aguaro, D. unpublished observation.
    2. Ellard, J., Greci, J. and Davis, T.B. unpublished observation.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?