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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-photo

    4 μg of ssRNA Ladder (denatured) on a 1.2% agarose-TBE 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 bases
    27,000 bases
    35,000 bases
    43,000 bases
    52,000 bases
    61,000 bases
    7500 bases

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM sodium citrate
    1 mM EDTA
    pH 6.0 @ 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 Cook, S. and Marchetti, C. Unpublished observation

    FAQs

    1. I need to know what types of gel to use to run RNA ladders?
    2. What types of gel and staining dyes can I use for NEB’s ssRNA Ladder (N0362S)?
    3. How much of the ssRNA Ladder do I need to load on a FlashGel 1.2% (Lonza)?

    Tech Tips


    Protocols

    1. Sample preparation using provided RNA Loading Dye (2X) (N0362)

    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.

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    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.