Amplifying whole insect genomes with multiple displacement amplification.
Many applications in insect genetics require repeated analyses on individual organisms. These include population genetic and genomics, linkage mapping, molecular systematics and map-based positional cloning. However, using the polymerase chain reaction, a limited number of analyses are possible with DNA isolated from whole bodies or parts of small or preserved specimens. We describe the optimization of a new technique, Multiple Displacement Amplification, for 100-400-fold amplification of whole mosquito genomes. We demonstrate that MDA amplifies genomic DNA directly from an adult leg or from organisms as small as a first instar mosquito larva. Genetic polymorphisms revealed at individual loci are the same whether using the original genomic DNA or MDA DNA as a template.
Gorrochotegui-Escalante
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Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA. normago@lamar.colostate.edu
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D000330:Aedes / Q000235:genetics*; D000818:Animals; D004247:DNA / Q000032:analysis / Q000235:genetics; D005260:Female; D017344:Genes, Insect / Q000235:genetics*; D005821:Genetic Techniques; D016678:Genome; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational
D004247:DNA