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  • Low Range PFG Marker

    Description

    A mixture of lambda DNA-HindIII fragments and lambda concatemers embedded in 1% LMP agarose and supplied in a GelSyringe™ dispenser. The lambda DNA monomer (48.5 kb) can be used as a reference point as its concentration has been adjusted to make it the brightest band in the pattern. Designed to be used as size markers for pulsed-field gel electrophoresis (PFG). Size range: 0.1-200 kb.

    N0350_thumb

    1% agarose gel, 6 V/cm, 15°C for 15 hours. Switch times ramped from 1-12 seconds.

    Properties and Usage

    Bases

    Fragmentbp
    12194000
    11145500
    1097000
    948500
    823100
    79420
    66550
    54360
    42320
    32030
    2560
    1130

    Effective Size Range

    130bp to 194,000bp

    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 Low Range Markers using a CHEF apparatus. The 1% agarose gel was run at 6 volts/cm using ramped pulse times from 1 to 12 seconds for 15 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. Bands larger than 194 kb are successively larger concatemers of Lambda DNA.
    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. Recommended plug size is 20 µl (two small graduations on the GelSyringe volume scale) which contains approximately 0.5 µg of DNA. Each GelSyringe yields 25+ plugs.
    5. Melting plugs will cause denaturation of concatemers.
    6. 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.

    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?