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  • 1 kb DNA Ladder

    Description

    A number of proprietary plasmids are digested to completion with appropriate restriction enzymes to yield 10 bands suitable for use as molecular weight standards for agarose gel electrophoresis. The digested DNA includes fragments ranging from 0.5-10.0 kilobases (kb). The 3.0 kb fragment has increased intensity to serve as a reference band. The approximate mass of DNA in each of the bands is provided (assuming a 0.5 μg load) for approximating the mass of DNA in comparably intense samples of similar size.

    Comes supplied with 1 vial of Gel Loading Dye, Purple (6X), no SDS.



    N3232_thumb

    1 kb DNA Ladder visualized by ethidium bromide staining on a 0.8% TAE agarose gel. Mass values are for 0.5 µg/lane.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    Gel Loading Dye, Purple (6X), no SDS256X

    Properties and Usage

    Bases

    FragmentMassbp
    14210,002
    2428,001
    3506,001
    4425,001
    5334,001
    61253,001
    7482,000
    8361,500
    9421,000
    10a21517
    10b21500

    Effective Size Range

    500bp 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 μg of 1 kb DNA Ladder diluted in sample buffer.
    3. All fragments have a 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. 1 kb 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, Purple (6X), no SDS:
      2.5% Ficoll®-400 
      10mM EDTA 
      3.3mM Tris-HCL (pH 8.0@25°C)
      0.02% Dye 1 
      0.001% Dye 2 
    7. When DNA ladders are run on a polyacrylamide gel, some reference bands may separate due to nucleic acid composition

    References

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

    FAQs

    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 Midori Green with the DNA Ladders from NEB?
    7. Can I use SYBR® with the DNA Ladders from NEB?

    Tech Tips

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

    Protocols

    1. End-labeling Protocol
    2. Suggested protocol for loading a DNA Ladder/marker

    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.

    Selection Tools

    Certificate of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

    Safety Data Sheet

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    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.