Culley Lab Protocols

General Information

For those just beginning to use isozymes, we recommend reading the following:

Kephart, S.R. 1990. Starch gel electrophoresis of plant isozymes: A comparative analysis of techniques. American Journal of Botany 77(5): 693-712.

Isozyme Staining Recipes - most of my recipes (given in the above protocol) are adapted from the following sources:

Acquaah, G. 1992. Practical Protein Electrophoresis for Genetic Research. Dioscorides Press, Portland, OR, USA. [great explanation of the electrophoretic and isozyme staining processes]

Morden, C.W., J. Doebley, and K.F. Schertz. 1987. A Manual of Techniques for Starch Gel Electrophoresis of Sorghum Isozymes. MP-1635. The Texas Agricultural Experiment Station, College Station, TX, USA. [ideal grinding buffer for Viola]

Soltis, D.E., C.H. Haufler, D.C. Darrow, and G.J. Gastony. 1983. Starch gel electrophoresis of ferns: A compilation of grinding buffers, gel and electrode buffers, and staining schedules. American Fern Journal 73(1): 9-27.

Wendel, J.F. and N.F. Weeden. 1989. Visualization and interpretation of plant isozymes. In: Isozymes in Plant Biology, D.E. Soltis and P.S. Soltis, eds. Dioscorides Press, Portland, OR, USA. [my most often-used set of staining recipes]

Werth, C.R. 1985. Implementing an isozyme laboratory at a field station. Virginia Journal of Science 36(1): 53-76. [ideal for Asclepias]

DNA Extraction:
		Standard Mini-Prep CTAB Technique Genogrinder 96-Well Plate Extraction (Coming Soon!)

Microsatellites:
		Microsatelllite Development Protocol - see Travis Glenn's website Multiplex PCR Protocol using the Qiagen Kit Dual Primer Labeling & Multiplex PCR Protocol Rick's Pollen PCR Protocol Fragment Analysis is done at Cornell University's Biotechnology Resource Center Genemapper Instructions [we now use Genemarker]

Inter-Simple Sequence Repeats (ISSRs):
		ISSR General Information PCR Protocol and Gel Instructions Gel Analysis and Generation of Data Data Analysis Trouble-shooting - If you need further help, visit Andi Wolfe's ISSR Resource Website, including a page detailing my primer optimization work.

Isozymes:
		Isozyme General Information Protocol for Grinding, Gel Electrophoresis, & Staining