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  • ssRNA Ladder

    Description

    The ssRNA Ladder is a set of 7 RNA molecules produced by in vitro transcription of a mixture of 7 linear DNA templates. The ladder sizes are: 9000, 7000, 5000, 3000, 2000, 1000 and 500 bases. The 3000 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 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 (see other side) problems associated with toxic chemicals and the difficulties encountered when staining and blotting formaldehyde gels.

    Sample Preparation
    The ssRNA Ladder is also compatible with formaldehyde-based loading buffers.

    N0362

    1 μg of ssRNA Ladder (denatured) on a 1.2% TBE agarose gel.

    Reagents Supplied

    The following reagents are supplied with this product:

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

    Properties and Usage

    Bases

    Fragmentbp
    19,000
    27,000
    35,000
    43,000
    52,000
    61,000
    7500

    Effective Size Range

    500bp to 9,000bp

    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.
    2. 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.

    References

    1. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, (2nd ed.).
    2. Liu, Y.-C. and Chou, Y.-C. (1990). Biotechniques. 9
    3. Cook, S. and Marchetti, C. Unpublished observation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. Can I use SYBR® with the DNA Ladders from NEB?
    1. Sample preparation using provided RNA Loading Dye (2X) (N0362)
    2. Sample preparation using denaturants for precise sizing

    To 5 ‘end label with T4 Polynucleotide Kinase, first dephosphorylate to remove 5’ triphosphates that are present as a result of invitro transcription. (for ssRNA, dsRNA,and Low Range ssRNA Ladders)