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Whole Genome Amplification for Sequencing and Applications in Conservation Genetics |
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| Authors: | JAN E. JANEČKA LON I. GRASSMAN JR. RODNEY L. HONEYCUTT MICHAEL E. TEWES |
| Affiliation: | 1. Feline Research Program, Caesar Kleberg Wildlife Research Institute, Texas A&M University-Kingsville, Kingsville, TX 78363, USA;2. Pepperdine University, Natural Sciences Division, Malibu, CA 90263, USA |
| Abstract: | Abstract: 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. |
| Keywords: | Convention on International Trade in Endangered Species of Wild Fauna and Flora endangered species mitochondrial DNA polymerase chain reaction whole genome amplification |