Fehlberg, Shannon , Tew, Jessica , Church, Kathleen .
Amplification and sequencing of genomic DNA extracted from cactus spines.
Isolation of DNA from cactus tissues for the purposes of gene amplification, sequencing, and genotyping typically presents problems in part due to the polysaccharide-based mucilage and secondary metabolites present in stem, fruit, and floral tissues. Additionally, large-scale sampling of cactus tissues in the field presents further challenges with respect to collection, storage, and transport of these tissues. The possibility of simplifying both procedures by collecting and extracting DNA from cactus spines had not previously been investigated. We reasoned that such a technique would be especially useful for investigators interested in population and conservation genetics of cactus species. As a preliminary step for studying the conservation genetics of the Federally endangered Arizona hedgehog cactus, Echinocereus arizonicus, we developed and refined techniques for the isolation of DNA from cactus spines. We used this technique to isolate DNA from 18 species of cacti from seven genera representing three subfamilies. Cactus spines were harvested from the living collection housed at the Desert Botanical Garden in Phoenix, Arizona. Several PCR protocols and primer pairs drawn from the literature for one nuclear and three plastid DNA regions were tested and modified to maximize results. We successfully extracted, amplified, and sequenced DNA from cactus spines for all species tested. We believe this protocol will be a useful option for those interested in studying the genetics of cacti.
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1 - Desert Botanical Garden, 1201 N. Galvin Pkwy., Phoenix, AZ, 85008, USA
Presentation Type: Oral Paper:Papers for Sections
Location: Delaware C/Hyatt
Date: Wednesday, July 11th, 2012
Time: 2:00 PM