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Journal of the American Pomological Society
(J Am Pom Soc)

American Pomological Society

Volume 65 Number 4 Article 4 Pages: 201-206
Year 2011 Month 10
Title: DNA Extraction Protocols from Dormant Buds of Twelve Woody Plant Genera
Authors: B. Gilmore, N. Bassil and K. Hummer
Citation
Abstract:
Standard plant DNA extraction protocols call for samples of newly expanding leaves and shoots yet analysis is sometimes needed when plants are dormant. We evaluated three DNA extraction protocols using dormant buds from 40 species and four hybrids of 12 genera. Two protocols were from ready-to-use kits (the Omega E-Z 96 Plant DNA Kit and the Fast ID 96-Well Genomic DNA Extraction Kit) and the third included commercial lysis and protein precipitation reagents (Qiagen). The genera included: Actinidia (Hardy Kiwi), Rubus (red raspberry), Ribes (gooseberry and currant), Cydonia (quince), Sorbus (mountain ash), Juglans (butternut), Amelanchier (service berry), Pyrus (pear), Mespilus (medlar), Corylus (hazelnut), Paeonia (peony), and Vaccinium (blueberry). In each of the genera tested, except for Juglans, both the Qiagen and Omega protocols generated large amounts of DNA (averaging 40 and 14.8 μg, respectively, from 30 to 36 mg of tissue) from dormant buds. For Juglans, none of these procedures provided satisfactory amounts of DNA from dormant buds. The positive result for 11 genera expanded the options for the sources of tissue as well as time of tissue collection for DNA extraction. The highest DNA yield was obtained with the Qiagen protocol, which was the least expensive of the three. However, in this protocol the bud scales must be removed to obtain a clear DNA extract. The Omega protocol may be more efficient if DNA is to be extracted from a large number of samples. In each of these 11 genera, DNA produced by at least one of the three protocols was of sufficient quality to apply in downstream molecular techniques, such as sequencing.

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