A DNA fingerprinting procedure for ultra high-throughput genetic analysis of insects

Schlipalius, D. I., Waldron, J., Carroll, B. J., Collins, P. J. and Ebert, P. R. (2001) A DNA fingerprinting procedure for ultra high-throughput genetic analysis of insects. Insect Molecular Biology, 10 6: 579-585.


Author Schlipalius, D. I.
Waldron, J.
Carroll, B. J.
Collins, P. J.
Ebert, P. R.
Title A DNA fingerprinting procedure for ultra high-throughput genetic analysis of insects
Journal name Insect Molecular Biology   Check publisher's open access policy
Publication date 2001
Sub-type Article
DOI 10.1046/j.0962-1075.2001.00297.x
Volume number 10
Issue number 6
ISSN 0962-1075
Start page 579
End page 585
Total pages 7
Place of publication Oxford, United Kingdom
Publisher Blackwell Science
Collection year 2001
Language eng
Subject C1
270207 Quantitative Genetics
620100 Field Crops
Abstract Existing procedures for the generation of polymorphic DNA markers are not optimal for insect studies in which the organisms are often tiny and background molecular Information is often non-existent. We have used a new high throughput DNA marker generation protocol called randomly amplified DNA fingerprints (RAF) to analyse the genetic variability In three separate strains of the stored grain pest, Rhyzopertha dominica. This protocol is quick, robust and reliable even though it requires minimal sample preparation, minute amounts of DNA and no prior molecular analysis of the organism. Arbitrarily selected oligonucleotide primers routinely produced similar to 50 scoreable polymorphic DNA markers, between individuals of three Independent field isolates of R. dominica. Multivariate cluster analysis using forty-nine arbitrarily selected polymorphisms generated from a single primer reliably separated individuals into three clades corresponding to their geographical origin. The resulting clades were quite distinct, with an average genetic difference of 37.5 +/- 6.0% between clades and of 21.0 +/- 7.1% between individuals within clades. As a prelude to future gene mapping efforts, we have also assessed the performance of RAF under conditions commonly used in gene mapping. In this analysis, fingerprints from pooled DNA samples accurately and reproducibly reflected RAF profiles obtained from Individual DNA samples that had been combined to create the bulked samples.
Keyword Biochemistry & Molecular Biology
Entomology
Rhyzopertha Dominica
Molecular Markers
Phosphine
Resistance
Genetic Mapping
Arbitrary Primers
Linkage Map
Amplification
Markers
Pcr
Polymorphisms
Resistance
Q-Index Code C1

Document type: Journal Article
Sub-type: Article
Collection: School of Biological Sciences Publications
 
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Created: Tue, 14 Aug 2007, 15:54:39 EST