Quick blots and nonradioactive detection of DNA probes for the identification of mosquitoes.
The quick blot protocol is an improved technique for preparing crude insect homogenates for hybridization to nucleic acid probes. Individual insects are ground in wells of a microtiter plate and transferred to a dot blot manifold. This allows preparation of multiple filters and provides uniformity and an orderly arrangement of samples. The high background detection resulting from use of crude insect homogenates with nonradioactive detection systems was eliminated by incubating quick blot filters in a laundry stain remover containing proteases. We used mosquito species-specific DNA probes to demonstrate the effectiveness of nonradioactive DNA labeling systems with quick blots.
Johnson
D W
DW
Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611.
Cockburn
A F
AF
Seawright
J A
JA
D000818:Animals; D009033:Culicidae / Q000235:genetics*; D015342:DNA Probes / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D009710:Nucleotide Mapping / Q000379:methods*; D011849:Radioactive Tracers; D013045:Species Specificity
D015342:DNA Probes; D011849:Radioactive Tracers