Amplifying whole insect genomes with multiple displacement amplification

被引:28
作者
Gorrochotegui-Escalante, N [1 ]
Black, WC [1 ]
机构
[1] Colorado State Univ, Dept Microbiol Immunol & Pathol, Ft Collins, CO 80523 USA
关键词
whole genome amplification; multiple displacement amplification; genomics; Aedes aegypti; SSCP;
D O I
10.1046/j.1365-2583.2003.00401.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:195 / 200
页数:6
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