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  • λ DNA-HindIII Digest

    Description

    The HindIII digest of lambda DNA (cI857ind1 Sam 7) yields 8 fragments suitable for use as molecular weight standards for agarose gel electrophoresis (1). 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, Purple (6X), no SDS



    N3012_thumb

    Lambda DNA-HindIII Digest visualized by ethidium bromide staining. 1.0% agarose gel.
    The full list of DNA fragments can be found in the chart below.

    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
    147723,130
    21949,416
    31356,557
    4904,361
    5482,322
    6422,027
    712564
    83125

    Effective Size Range

    125bp to 23,130bp

    Storage Temperature

    -20°C

    Storage Conditions

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

    Notes

    1. 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 and subsequently heated. Temperatures >60°C may cause denaturation.
    2. The cohesive ends of fragments 1 and 4 may be separated by heating to 60°C for 3 minutes.
    3. 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

    References

    1. Daniels, D.L. et al. (1983). Appendix II: Complete Annotated Lambda Sequence. R.W. Hendrix, J.W. Roberts, F.W. Stahl and R.A. Weisberg(Ed.), Lambda-II. 519-676. New York: Cold Spring Harbor Laboratory Press.
    2. Forster, A.C. et al. (1985). Nucl. Acids Res. 13, 745-761.

    FAQs

    1. 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?
    2. How can I quantify the amount of DNA in each band of a marker?
    3. How do I use the lambda DNA-Hind III digest?
    4. Why does my Lambda DNA molecular weight marker appear smeared and not represent the correct banding pattern?
    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?

    Protocols

    1. Suggested protocol for loading a sample

    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.

    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.