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  • Low Molecular Weight DNA Ladder

    Description

    A proprietary plasmid is digested to completion with appropriate restriction enzymes to yield 11 bands suitable for use as molecular weight standards for both agarose and polyacrylamide gel electrophoresis. This digested DNA includes fragments ranging from 25-766 base pairs. The 200 base pair band has increased intensity to serve as a reference point.

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



    N3233-new-photo

    LMW DNA Ladder visualized by ethidium bromide staining on a 3% TBE 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
    142766
    227500
    320350
    433300
    527250
    6110200
    733150
    843100
    95875
    106350
    114325

    Effective Size Range

    25bp to 766bp

    Storage Temperature

    -20°C

    Storage Conditions

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

    Notes

    1. We recommend loading 0.5 µg of the Low Molecular Weight DNA Ladder diluted in sample buffer.
    2. 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.
    3. All ends have 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] dCTP or α-[32P] dGTP for the fill-in reaction.
    4. DNA ladders are 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 dH2O
    6. 1X Gel Loading Dye, Purple, 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. Due to the limitations of the acrylamide gel technology, one or two extra bands may be visible on the DNA ladders when run on a polyacrylamide gel.

    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. Why is the separation of the lower bands incomplete?
    3. 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?
    4. Why are the DNA ladders showing up on my Southern blot? What is the sequence or composition of the ladder bands?
    5. How can I quantify the amount of DNA in each band of a marker?
    6. Can I use GelRed with the DNA Ladders from NEB?
    7. Can I use Midori Green with the DNA Ladders from NEB?
    8. 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

    Interactive 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]

    Specifications

    The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

    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.