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  • 2-Log DNA Ladder (0.1-10.0 kb)

    Description

    A number of proprietary plasmids are digested to completion with appropriate restriction enzymes to yield 19 bands suitable for use as molecular weight standards for agarose gel electrophoresis. This digested DNA includes fragments ranging from 100 bp to 10 kb. The 0.5, 1.0 and 3.0 kb bands have increased intensity to serve as reference points. The approximate mass of DNA in each of the bands is provided (assuming a 1.0 μg load) for approximating the mass of DNA in comparably intense samples of similar size.

    Comes supplied with 1 vial of Gel Loading Dye, Blue (6X).



    N3200_thumb

    2-Log DNA Ladder visualized by ethidium bromide staining on a 1.0% TBE agarose gel. Mass values are for 1 µg/lane.

    Properties and Usage

    Bases

    FragmentMassbp
    14010002
    2408001
    3486001
    4405001
    5324001
    61203001
    7402017
    8571517
    9451200
    101221000
    1134900
    1231800
    1327700
    1423600
    15124500/517
    1649400
    1737300
    1832200
    1961100

    Effective Size Range

    100bp to 10,002bp

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    1 mM EDTA
    pH 8.0 @ 25°C

    Notes

    1. This ladder was not designed for precise quantification of DNA mass but can be used for approximating the mass of DNA in comparably intense samples of similar size.
    2. We recommend loading 0.5-1 μg of the 2-Log DNA Ladder diluted in sample buffer.
    3. All fragments have 4-base, 5´ overhangs that can be end labeled using T4 Polynucleotide Kinase (#M0201) or filled-in using DNA Polymerase I, Klenow Fragment (#M0210) (1). Use α-[32P] dATP or α-[32P] dTTP for the fill-in reaction.
    4. 2-Log DNA Ladder is stable for at least 3 months at 4°C.
    5. For long term storage, store at -20°C. If samples need to be diluted, use TE or other buffer of minimal ionic strength. DNA may denature if diluted in dH20
    6. 1X Gel Loading Dye, Blue:
      2.5% Ficoll-400
      11 mM EDTA
      3.3 mM Tris-HCl (pH 8.0@25°C)
      0.017% SDS
      0.015% bromophenol blue
    7. When DNA ladders are run on an acrylamide gel, some separation of the reference bands may be observed due to variations in the nucleic acid composition of the DNA molecules within that band.

    References

    1. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (2nd Ed.). 10.51-10.67.

    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. Why are there extra bands visible on polyacrylamide gels?
    2. What are the overhangs on the DNA ladder fragments? Can I end-label them using the T4 polynucleotide kinase (PNK)? What about the Klenow fragment?
    3. Why are the DNA ladders showing up on my Southern blot? What is the sequence or composition of the ladder bands?
    4. How can I quantify the amount of DNA in each band of a marker?
    5. Can I use GelRed with the DNA Ladders from NEB?
    6. Can I use SYBR® with the DNA Ladders from NEB?
    7. Can I use Midori Green with the DNA Ladders from NEB?
    1. End-labeling Protocol
    2. Suggested protocol for loading a sample (N3200)
    3. Suggested protocol for loading a sample

    Selection Tools

    To make it ready-to-load, dilute in TE buffer instead of water.