Rapid DNA extraction methods and new primers for randomly amplified polymorphic DNA analysis of Giardia duodenalis.
A randomly amplified polymorphic DNA (RAPD) procedure using simple genomic DNA preparation methods and newly designed primers was optimized for analyzing Giardia duodenalis strains. Genomic DNA was extracted from in vitro cultivated trophozoites by five freezing-thawing cycles or by sonic treatment. Compared to a conventional method involving proteinase K digestion and phenol extraction, both freezing-thawing and sonication were equally efficient, yet with the advantage of being much less time- and labor-intensive. Five of the 10 tested RAPD primers produced reproducible polymorphisms among five human origin G. duodenalis strains, and grouping of these strains based on RAPD profiles was in agreement among these primers. The consistent classification of two standard laboratory reference strains, Portland-1 and WB, in the same group confirmed previous results using other fingerprinting methods, indicating that the reported simple DNA extraction methods and the selected primers are useful in RAPD for molecular characterization of G. duodenalis strains.
Deng
M Q
MQ
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California at Davis, 95616-8743, USA.
Cliver
D O
DO
D000818:Animals; D017931:DNA Primers; D016054:DNA, Protozoan / Q000302:isolation & purification*; D005872:Giardia / Q000235:genetics*; D006801:Humans; D019105:Random Amplified Polymorphic DNA Technique
D017931:DNA Primers; D016054:DNA, Protozoan