Whole Genome Amplification for Sequencing and Applications in Conservation Genetics

• Published in: The Journal of wildlife management Vol. 71; no. 4; pp. 1357 - 1360
• Main Authors: JANEČKA, JAN E, GRASSMAN, LON I, HONEYCUTT, RODNEY L, TEWES, MICHAEL E
• Format: Journal Article
• Language: English
• Published: Oxford, UK The Wildlife Society 01-06-2007; Blackwell Publishing Ltd
• Subjects: Animal genetics; animal tissues; Cats; Conservation genetics; conservation programs; Convention on International Trade in Endangered Species of Wild Fauna and Flora; Deoxyribonucleic acid; DNA; DNA polymerase; DNA polymerase-mediated strand displacement reaction; duration; Ecological genetics; endangered species; Genetic markers; Genetic research; Genetics; genome; Genomes; Genomics; Leopardus pardalis; Methods; Mitochondrial DNA; Neofelis nebulosa; Polymerase chain reaction; Prionailurus bengalensis; R&D; Research & development; Research methods; sequence analysis; Studies; Techniques and Technology; Techniques and Technology Articles; whole genome amplification; wild animals; Wildlife conservation; zoo animals
• ISSN: 0022-541X; 1937-2817
• DOI: 10.2193/2006-369

We describe a method for rapidly amplifying whole genomes via a Phi29 DNA polymerase-mediated strand displacement reaction (SDR). Genomic amplification products derived from the SDR reaction resulted in high quantities of DNA suitable for polymerase chain reaction (PCR) amplification and sequencing of mitochondrial genomes. Control region sequences of DNA derived directly from PCR amplicons of extracted DNA were identical to those derived from PCR amplification of SDR genomic DNA. Effective SDR amplification and subsequent sequencing was successful across tissues sources ranging in age from 1 year to 19 years. Strand replacement reaction genomic amplification offers a means of obtaining large quantities of DNA from small amounts of tissue.