Penstemon clevelandii
Why the pictures of these three taxa? The diploid hybrid species status of P. clevelandii was documented with ISSR markers (Wolfe et al. 1998a, b) and we've used ISSR markers successfully to examine relationships within the holoparasitic plant genus Hyobanche and the conservation genetics of P. debilis.
Department of Evolution, Ecology and Organismal Biology
The Ohio State University
1735 Neil Avenue
Columbus OH 43210
Bibliography (ISSR
citations, other references of interest, some notes on selected
citations)
Data Gathering and Scoring (general
considerations, types of data needed for selected studies)
Introduction (overview of this
website, history of method)
Protocols: DNA Miniprep extraction
Protocols: Elu-Quik clean-up of
DNA extract
Protocols: ISSR primers and PCR(NEW
primers listed as of 1/17/00)
Protocols: ISSR primer optimization
and troubleshooting (
All photos labeled as of 1/17/00)
Protocols: Sephaglas clean-up
of DNA extract
Resources (links and addresses for
useful sites, including software)
Software and Algorithms (so many
choices, so little memory)
What's New (a chronology of ISSR Resource
Website development)
19 July 2000
Updated the bibliography - new citations include ISSR references for insect, mammal, algae, and fungi publications.
17 January 2000
New primers are listed and the opitimization page now includes the labeled gel pics.
29 October 1999
I've added a page on optimization and trouble shooting ISSR gels. This page includes pictures and explanations of each step.
17 November 1998
I've added a Sephaglas protocol, a page on resources, and a page on software and algorithms. The latter page includes examples of input and output for Vera S. Ford's programs, which we have cited in several manuscripts. I've included instructions for obtaining these programs through anonymous FTP.
Except for adding examples from case studies and a FAQ page, this website includes the relevant information I included in my 1998 workshop at AIBS. I'll continue to update the bibliography on a regular basis, and I will start developing the add-ons as I have time (oh sure, like I'm really going to have time).
19 September 1998
I've added the Elu-Quik protocol we use.
24 August 1998
There's a section for data gathering/scoring.
20 August 1998
I've added miniprep DNA extraction and ISSR protocols.
Send me feedback! via this link or by leaving a message in the guestbook (click on the Guest book button of the banner below).
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The first studies employing ISSR markers were published in 1994 (Zietkiewicz et al. 1994; Gupta et al. 1994). The initial studies focussed on cultivated species, and demonstrated the hypervariable nature of ISSR markers (reviewed in Wolfe and Liston 1998). To test the utility of the method in natural populations, Wolfe et al. (1998a, b) reexamined a known hybrid complex of four species of Penstemon for which three other molecular data sets were available. Their results clearly demonstrated the utility of ISSR markers for addressing questions of hybridization and diploid hybrid speciation. In addition, several other studies from A. Liston's lab at Oregon State University, A. Wolfe's and D. Crawford's labs at Ohio State University and H. Ballard's lab at Ohio University ranging from conservation biology to molecular ecology and systematics have demonstrated the utility of the technique in a wide range of applications and plant families (Asteraceae, Brassicaceae, Hippocastanaceae, Orchidaceae, Poaceae, Scrophulariaceae, Violaceae).
ISSR techniques are nearly identical to RAPD techniques except that ISSR primer sequences are designed from microsatellite regions and the annealing temperatures used are higher than those used for RAPD markers. Based on the published, unpublished and in-progress studies that have been conducted using ISSR markers, it is clear that ISSR markers have great potential for studies of natural populations (reviewed in Wolfe et al. 1998b).
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Please send your suggestions, comments,
corrections to wolfe.205@osu.edu
Last updated January 17, 2000.
Wolfe Homepage
Wolfe Lab Homepage
Penstemon Website