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Plants 2017, 6(1), 1; doi:10.3390/plants6010001

A High-Throughput RNA Extraction for Sprouted Single-Seed Barley (Hordeum vulgare L.) Rich in Polysaccharides

1
United States Department of Agriculture—Agriculture Research Service (USDA-ARS), Small Grains and Potato Germplasm Research Unit, 1691 S. 2700 W., Aberdeen, ID 83210, USA
2
Department of Plant and Wild Life Sciences, Brigham Young University (BYU), Provo, UT 84602, USA
*
Authors to whom correspondence should be addressed.
Academic Editors: Vagner A. Benedito and Mark F. Belmonte
Received: 31 August 2016 / Revised: 9 December 2016 / Accepted: 20 December 2016 / Published: 22 December 2016
View Full-Text   |   Download PDF [2158 KB, uploaded 22 December 2016]   |  

Abstract

Germinated seed from cereal crops including barley (Hordeum vulgare L.) is an important tissue to extract RNA and analyze expression levels of genes that control aspects of germination. These tissues are rich in polysaccharides and most methods for RNA extraction are not suitable to handle the excess polysaccharides. Here, we compare the current methods for RNA extraction applicable to germinated barley tissue. We found that although some of these standard methods produced high-quality RNA, the process of extraction was drastically slow, mostly because the frozen seed tissue powder from liquid N2 grinding became recalcitrant to buffer mixing. Our suggested modifications to the protocols removed the need for liquid N2 grinding and significantly increased the output efficiency of RNA extraction. Our modified protocol has applications in other cereal tissues rich in polysaccharides, including oat. View Full-Text
Keywords: barley; cereals; sprouted seed; germination; high-quality RNA; single-seed extraction; qRT-PCR barley; cereals; sprouted seed; germination; high-quality RNA; single-seed extraction; qRT-PCR
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Rashid, A.; Baldwin, T.; Gines, M.; Bregitzer, P.; Esvelt Klos, K. A High-Throughput RNA Extraction for Sprouted Single-Seed Barley (Hordeum vulgare L.) Rich in Polysaccharides. Plants 2017, 6, 1.

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