| Abstract Detail
Recent Topics Posters Barrera, Daniel [1], Kates, Heather Rose [2], Soltis, Pamela S. [3], Soltis, Douglas [4]. Preparing nonviable Cucurbita seeds for next generation DNA sequencing to enable diverse collaborations in studies of crop wild relatives. Crop wild relatives (CWR) will play an important role in improving food security as crops adapt to climate change and population growth. Although the importance of studying CWR is clear, many CWR are threatened in the wild and CWR seeds available for research are sometimes non-viable due to poor storage conditions. Here we test the relative utility of four DNA-extraction-from-seed protocols for genomic-library-building and next-generation-sequencing (NGS) on a set of non-viable seeds from accessions of giant pumpkin (Cucurbita maxima ssp. maxima) and its CWR (Cucurbita maxima ssp. andreana) that are needed for a population genomics study to characterize the genetic consequences of domestication in giant pumpkin. We compared the quality and yield of DNA extracted in the samples across the four protocols and found that DNA extraction from ground embryonic seed tissue using a modified-CTAB protocol with two phenol purification steps provided the best results. We built and quantified genomic libraries for a subset of our resulting DNA samples to confirm that the DNA extracted from seed was usable for NGS applications.
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1 - Florida Museum of Natural History, Phylogenetics, 1659 Museum Rd, Gainesville, FL, 32611, USA
2 - University of Florida, Florida Museum of Natural History, Dickinson Hall, PO Box 117800, Gainesville, FL, 32611, USA
3 - University Of Florida, Florida Museum Of Natural History, PO BOX 117800, Gainesville, FL, 32611-7800, USA, 352/273-1964
4 - Florida Museum of Natural History, Florida Museum of Natural History, Dickinson Hall - Museum Road, Gainesville, FL, 32611, United States
Keywords:
crop wild relative
cucurbitaceae.
Presentation Type: Recent Topics Poster
Session: P, Recent Topics Posters
Location: Exhibit Hall/Savannah International Trade and Convention Center
Date: Monday, August 1st, 2016
Time: 5:30 PM
Number: PRT032
Abstract ID:1225
Candidate for Awards:None |
