Evaluation of three methods of extraction in quality of DNA in stool samples of deer-catingueiro (Mazama gouazoubira)

• Grant number: 12/08823-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2012
• End date: June 30, 2013
• Field of knowledge: Biological Sciences - Genetics
• Principal Investigator: José Maurício Barbanti Duarte
• Grantee: Aline Aparecida Silva Stella
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

The increasing fragmentation of habitats has led to a decrease in populations of Brazilian deer and consequently reduction of their genetic variability. Research to estimate this damage have been developed, allowing the establishment of management programs and conservation of these species. Thus, the use of non-invasive methods used since the 1990s, has proven to be an important tool in conservation genetics. Through them it has been possible to extract and amplify DNA from various biological matrices, generating knowledge about the population density, habitat use, species identification, sexing, among others. One limiting the use of this type of sampling is the inability to quantify the DNA from these samples in the spectrometry apparatus traditional. However, as the Polymerase Chain Reaction (PCR) in real time is possible to quantify the amplified DNA during each cycle of the reaction, allowing to monitor the quantity and therefore the quality of each fecal sample, which minimizes the high financial cost and time employed in the process of re-amplified samples for the target region. In this study, fecal samples of Mazama gouazoubira will be extracted from three different methods available in the literature for this type of non-invasive sampling. From a pair of primers for heterologous region microsatellite and a pair of primers to the region of cytochrome B (CytB) is checked whether there influence of extract used, as well as the quality of the samples (determined at the time of collection) from the measurement of DNA amplifiable by these primers in PCR in real time. (AU)