TITLE: RNA Isolation from Plant Tissue
AUTHORS: Marc T. J. Johnson, Eric J. Carpenter, Zhijian Tian, Richard Bruskiewich, Jason N. Burris, Charlotte T. Carrigan, Mark W. Chase, Neil D. Clarke, Sarah Covshoff, Claude W. dePamphilis, Patrick P. Edger, Falicia Goh, Sean Graham, Stephan Greiner, Julian M. Hibberd, Ingrid Jordon-Thaden, Toni M. Kutchan, James Leebens-Mack, Michael Melkonian, Nicholas Miles, Henrietta Myburg, Jordan Patterson, J. Chris Pires, Paula Ralph, Megan Rolf, Rowan F. Sage, Douglas Soltis, Pamela Soltis, Dennis Stevenson, C. Neal Stewart Jr, Barbara Surek, Christina J. M. Thomsen, Juan Carlos Villarreal, Xiaolei Wu, Yong Zhang, Michael K. Deyholos, Gane Ka-Shu Wong

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Methods for RNA isolation</span></div><div class = "text-block"><span>These methods were originally included in Appendix S1 of "Evaluating Methods for  Isolating Total RNA and Predicting the Success of Sequencing  Phylogenetically Diverse Plant Transcriptomes" Marc T. J. Johnson et al.  PLOS ONE, November 21, 2012.  </span><a href="https://doi.org/10.1371/journal.pone.0050226" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">https://doi.org/10.1371/journal.pone.0050226</span></a></div><div class = "text-block">Many of the protocols share elements or combine components from several methods. For each method, we describe the reagents and procedures used, and identify the researchers or institute that implemented the protocol.</div><div class = "text-block">Due to the potential for contamination and degradation by RNase enzymes, as well as health concerns in handling some substances and chemicals, best practices in aseptic wet lab techniques must be practiced at all times during RNA isolation. Chief among these are the critical need to avoid contamination of samples by using extreme care when moving liquids and opening and closing tubes to avoid aerosols. Because of the risk of degradation by RNase enzymes, it is essential to use sterile RNase-free equipment, disposable plastics and solutions.</div><div class = "text-block"><span>RNase degradation and contamination can be avoided by keeping samples constantly frozen at low temperature (< -80 °C) prior to adding buffers that denature or immobilize RNase. Treating equipment with RNase denaturants (e.g. RNase Zap, Ambion, Austin, TX) and solutions with diethylpyrocarbonate (DEPC) can also prevent contamination and/or degradation of samples, but it can have some negative effects on samples</span><span style = "vertical-align:super;">1</span><span>. Many additional helpful tips for successful RNA isolation are available in Sambrook and Russell</span><span style = "vertical-align:super;">1</span><span> and in Appendix A of </span><a href="https://www.qiagen.com/es/resources/resourcedetail?id=14e7cf6e-521a-4cf7-8cbc-bf9f6fa33e24&lang=en" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">Qiagen’s RNeasy Mini Handbook</span></a><span> downloadable from </span><a href="https://www.qiagen.com/es/" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">www.qiagen.com</span></a><span>.</span></div><div class = "text-block"><span style = "vertical-align:super;">1</span><span> Sambrook, J. & Russell, D.W. Molecular Cloning: A Laboratory Manual, 3rd ed. (Cold Spring Harbor</span></div><div class = "text-block">Laboratory Press, Cold Spring Harbor, NY, 2001).</div></div>

[STEPS]