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  • PCR Marker

    Description

    The PCR Marker consists of a proprietary plasmid that is digested to completion with appropriate restriction enzymes to yield 5 double-stranded DNA bands suitable for use as molecular weight standards for agarose and acrylamide gel electrophoresis. The digested DNA includes fragments ranging from 50-766 base pairs. The approximate mass of DNA in each of the bands is provided (assuming a 0.3 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.




    N3234_thumb

    PCR Marker visualized by ethidium bromide staining on a 1.8% TBE Agarose Gel. Mass values are for 0.3 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
    162766
    240500
    348300
    461150
    58950

    Effective Size Range

    50bp to 766bp

    Storage Temperature

    4°C

    Storage Conditions

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

    Notes

    1. We recommend loading 0.3 μg of PCR Marker diluted in sample buffer.
      This marker 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. PCR Marker is stable for at least 3 months at 4°C. 
    3. 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.
    4. All ends have 5' overhangs that can be labeled using T4 Polynucleotide Kinase (NEB #M0201) or filled-in using DNA Polymerase I, Klenow Fragment (NEB #M0210) (1). Use α-[32P] dCTP or α-[32P] dGTP for the fill-in reaction.
    5. 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

    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. Can I use GelRed with the DNA Ladders from NEB?
    4. Can I use Midori Green with the DNA Ladders from NEB?
    5. Can I use SYBR® with the DNA Ladders from NEB?
    6. Why are there extra bands visible on polyacrylamide gels?

    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 sample (PCR 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]

    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.