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  • Low Range ssRNA Ladder

    Description

    The Low Range ssRNA Ladder is a set of 6 RNA molecules produced by in vitro transcription of a mixture of 6 linear DNA templates. The ladder sizes are: 1000, 500, 300, 150, 80 and 50 bases. The 300 base fragment is at double intensity to serve as a reference band. This ladder is suitable for use as an ssRNA size standard on denaturing polyacrylamide-urea gels, and on denaturing or native agarose gels.

    Usage Recommendation
    This marker was not designed for precise quantification of ssRNA mass.

    Denaturing vs. Native Agarose Gels
    It is common practice to electrophorese RNA on a fully denaturing agarose gel, such as one containing formaldehyde (1). However, in many cases it is possible to run RNA on a native agarose gel and obtain suitable results. In fact, it has been demonstrated that treatment of RNA samples in a denaturing buffer maintains the RNA molecules in a denatured state, during electrophoresis, for at least 3 hours (2,3). The use of native agarose gels eliminates problems associated with toxic chemicals and the difficulties encountered when staining and blotting formaldehyde gels.

    Sample Preparation
    The Low Range ssRNA Ladder is also compatible with formaldehydebased loading buffers.

    N0364_thumb

    0.25 μg of Low Range ssRNA Ladder (denatured) on a 2.0% TBE agarose gel stained with

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    RNA Loading Dye, (2X)2X

    Properties and Usage

    Bases

    Fragmentbp
    11,000
    2500
    3300
    4150
    580
    650

    Effective Size Range

    50bp to 1,000bp

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM Potassium Acetate
    pH 4.5 @ 25°C

    Notes

    1. Minimize repeated freeze-thaw cycles. It is best to aliquot the marker into single use portions.

      To avoid ribonuclease contamination: wear gloves, use RNase-free water for gels and buffers, wash equipment with detergent and rinse thoroughly with RNase-free water.

    2. Store at -70°C. For short term storage (< 1 week), ladder can be stored at -20°C.

    References

    1. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. 2nd ed., 743-745.
    2. Liu, Y-C. and Chou, Y-C. (1990). Biotechniques. 9
    3. Sandra Cook, and Christina Marchetti Unpublished observation
    4. Dong Ma, and John Greci Unpublished observation

    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. How to prepare denatured RNA samples to be run on a native gel?
    2. Can I use SYBR® with the DNA Ladders from NEB?
    1. Sample preparation using provided buffer (N0364)
    2. Sample preparation using denaturants for precise sizing
    Only 0.25 ug /lane is needed for a denaturing polyacrylamide-urea gel.
    6% denaturing polyacrylamide-urea gels give the best resolution of all 6 bands.