College
College of Arts and Sciences
Mentor Information
Michelle Green
Description
The lowland anoa (Bubalus depressicornis) is also known as the dwarf water buffalo endemic to the Island of Sulawesi. This species is considered endangered under the IUCN Red List with less than 2,500 individuals remaining in the wild. Genetic research for anoa is insufficient because of population fragmentation and their avoidance of humans. Utilizing noninvasive samples alleviates the need for direct animal interaction and can provide DNA suitable for population level analysis of genetic diversity. DNA was previously isolated from anoa fecal and saliva samples. In this study, both fecal and saliva samples were tested, with focus on saliva samples. Saliva DNA was isolated via Quick-DNA MICROprep kit while fecal samples were isolated by Qiagen FAST DNA kit. To determine whether this DNA is sufficient, it must be amplified using a set of genetic markers. In this study, primer TGLA126 was selected for testing because of its success as a microsatellite marker in other bovine studies and in a study analyzing tissue samples of anoa. TGLA126 targets a microsatellite locus with an allele size range of 104-109 bps, making it suitable for PCR amplification. To determine the effectiveness of TGLA126 for the lowland anoa, multiple PCR trials were completed. Trials differed by annealing temperature (45-58°C), concentration of DNA and primer, and sample type. This study expects to find that primer TGLA126 is successful in DNA amplification of saliva and fecal samples. Implications of determining the viability of primer TGLA126 include helping future researchers approach conservation strategies of Bubalus depressicornis.
Evaluating TGLA126 for PCR Amplification of Noninvasive DNA Samples in the Lowland Anoa
The lowland anoa (Bubalus depressicornis) is also known as the dwarf water buffalo endemic to the Island of Sulawesi. This species is considered endangered under the IUCN Red List with less than 2,500 individuals remaining in the wild. Genetic research for anoa is insufficient because of population fragmentation and their avoidance of humans. Utilizing noninvasive samples alleviates the need for direct animal interaction and can provide DNA suitable for population level analysis of genetic diversity. DNA was previously isolated from anoa fecal and saliva samples. In this study, both fecal and saliva samples were tested, with focus on saliva samples. Saliva DNA was isolated via Quick-DNA MICROprep kit while fecal samples were isolated by Qiagen FAST DNA kit. To determine whether this DNA is sufficient, it must be amplified using a set of genetic markers. In this study, primer TGLA126 was selected for testing because of its success as a microsatellite marker in other bovine studies and in a study analyzing tissue samples of anoa. TGLA126 targets a microsatellite locus with an allele size range of 104-109 bps, making it suitable for PCR amplification. To determine the effectiveness of TGLA126 for the lowland anoa, multiple PCR trials were completed. Trials differed by annealing temperature (45-58°C), concentration of DNA and primer, and sample type. This study expects to find that primer TGLA126 is successful in DNA amplification of saliva and fecal samples. Implications of determining the viability of primer TGLA126 include helping future researchers approach conservation strategies of Bubalus depressicornis.
