Working with RNA

Set up a lab RNA Zone

Working with RNA can be a tricky thing…it falls apart easily, and RNases (enzymes that degrade RNA) are ubiquitous. Successfully isolating RNA and maintaining its integrity is critical, especially when sensitive downstream applications are used (e.g., RNA-Seq).

Good techniques for RNA handling are simple to employ but crucial for success. All RNA purification and handling should take place in an RNase-free, RNA-only zone of the lab. Segregating RNA work from protein and DNA purification and handling will help minimize the potential for RNase contamination and help keep your RNA intact. Only buffer and water stocks treated to be RNase-free should be kept in the RNA area of the lab, and gloves should be worn at all times to prevent accidental contamination. Tools and equipment such as pipets, tips, and centrifuges should be designated for use only in the RNA zone as well. The location of the RNA zone in the lab is also important. Keeping traffic to a minimum and moving the RNA zone away from doors, windows, and vents can also help minimize contamination.

Using an RNase inhibitorcan also help safeguard your samples from RNase degradation. These inhibitors can bind to any RNases that may have been introduced into your sample and prevent them from cutting the RNA present.

Water and buffer stocks can be a source of RNase contamination. Several stocks from an RNase-free zone in an academic lab showed RNase activity. Recombinant RNasin® inhibitor protected all RNA samples from degradation.

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3 thoughts on “Working with RNA

  1. Kind regards,
    Will you be able to guide how to make an exclusive RNA working lab – that one will be help. i.e. hand care while handling, working areas managements with RNase sprays – the compositions etc..,
    With regards,
    S. AFSAR

    • Dear S. Afsar,

      Thank you for your question. The following article, titled “Working with RNA” , explains in detail how to establish an RNase-free enviroment, which is crucial when working with RNA:
      http://www.promega.com/~/media/files/resources/product%20guides/rna%20analysis%20notebook/workingwithrna.pdf?la=en . In addition to these recommendations it is advice to clean working space, bench surfaces, pipettes, electrophoresis equipment, among other things with an RNase cleaning product (0.5%SDS, then rinse with RNase-free water, and then rinsed with ethanol and air dried).

      I hope that this information is useful. Please let me know if you have further questions.

      Best regards,

      Laura

      Laura A. Pérez-Casellas, Ph.D.
      Technical Services Scientist
      Promega Corporation
      Tel 1 800 356 9526 Ext 1440 (USA)
      techserv@promega.com

      • Thank you sir for your kind regards provided here.
        We are in general using the precautions as you mentioned and along with this if there lies any other suggestions updated from you people – try to let me know here.

        With regards,
        S. AFSAR

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