Filter paper for preservation, storage, and distribution of insect and pathogen DNA samples.
نویسندگان
چکیده
Proper DNA storage is critical for studies involving genetic analysis of insects and for molecular diagnostics of pathogens carried by them. Molecular surveillance of pathogens carried by insects can involve screening of thousands of insect DNA samples. Problems with storage and degradation of these samples can arise. In this study, a simple filter paper-based method for storage and preservation of insect DNA was evaluated using polymerase chain reaction (PCR). DNA was isolated from individual house fly, Musca domestica L., adults by using a cell lysis technique. From 50 house flies known to carry Campylobacter spp., a portion of the DNA sample was stored frozen and another portion was pipetted onto filter paper. At monthly intervals for 7 mo, PCR was conducted using 1 microl of the frozen DNA sample and a 2.0-mm disk from the filter paper samples as the PCR template. Two markers were used, a 450-bp region of the insect mitochondrial DNA (mtDNA) ND5 gene and a 857-bp region of the Campylobacter spp. mtDNA 16S rDNA gene. PCR amplification was successful for all of the samples regardless of the storage method. The filter paper method is a simple and economical way to store, preserve, and distribute DNA samples for PCR analysis.
منابع مشابه
Carrie B . Owens and Allen L . Szalanski University of Arkansas , Department of Entomology , Fayetteville , AR Filter paper for preserving insect and pathogen DNA samples
We thank M.J.Toliver and W.J. Hatfield for their technical assistance. This work was supported by the University of Arkansas, Arkansas Agricultural Experiment Station, and by USDA CSREES Pest Management Alternatives Grant 2002-34381 -12156 and USDA CSREES Food Safety Consortium Grant 2001 -34211-10288. Carrie B. Owens and Allen L. Szalanski University of Arkansas, Department of Entomology, Faye...
متن کاملUtility of Filter Paper for Preserving Insects, Bacteria, and Host Reservoir DNA for Molecular Testing
BACKGROUND Appropriate methodology for storage biological materials, extraction of DNA, and proper DNA preservation is vital for studies involving genetic analysis of insects, bacteria, and reservoir hosts as well as for molecular diagnostics of pathogens carried by vectors and reservoirs. Here we tried to evaluate the utility of a simple filter paper-based for storage of insects, bacteria, rod...
متن کاملInfluence of preserved brewing yeast strains on fermentation behavior and flocculation capacity
Preservation methods on the physiological and brewing technical characters in bottom and top brewing yeast strains were investigated. The preserved yeasts were reactivated after 24 months storage and grown up to stationary phase. The samples of filter paper storage indicated a higher cell growth and viability during propagation than those of nitrogen and lyophilization storage independent on pr...
متن کاملPaper-based archiving of mammalian and plant samples for RNA analysis.
The ability to archive biological samples for subsequent nucleic acid analysis is essential for tissue specimens and forensic samples. FTA Card is a chemically treated filter paper designed for the collection and room temperature storage of biological samples for subsequent DNA analysis. Its usefulness for the preservation of biological samples for subsequent RNA analysis was tested. Here, we d...
متن کاملThe room temperature preservation of filtered environmental DNA samples and assimilation into a phenol–chloroform–isoamyl alcohol DNA extraction
Current research targeting filtered macrobial environmental DNA (eDNA) often relies upon cold ambient temperatures at various stages, including the transport of water samples from the field to the laboratory and the storage of water and/or filtered samples in the laboratory. This poses practical limitations for field collections in locations where refrigeration and frozen storage is difficult o...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Journal of medical entomology
دوره 42 4 شماره
صفحات -
تاریخ انتشار 2005