College
College of Arts and Sciences
Mentor Information
Michelle Green
Description
The lowland anoa (Bubalus depressicornis) is the smallest species of wild cattle in the world and is classified as endangered on the ICUN Red List. Noninvasive genetic assessment allows for collection of samples with minimal impact on animal welfare but can result in low-quality DNA or contamination. Little is known about effective forms of noninvasive sampling and the DNA analysis protocol that follows regarding the anoa. This study is part of a continuing collaborative investigation and helped gain insight into the suitability of noninvasive DNA for population research. To determine whether the DNA is suitable, a set of genetic markers is needed. This study tested whether noninvasive DNA from fecal and saliva samples would amplify using primer CSRM60. CSRM60 was selected based on cross-species amplification in bovids, (e.g., cattle and domestic buffaloes). CSRM60 targets a microsatellite locus with an allele size range of 83-99 bp. This study primarily tested DNA taken from fecal samples stored in ethanol and extracted using a Qiagen Fast DNA Kit. Saliva samples tested were stored in DNA/RNA shield and extracted by Quick-DNA MICROprep Kit. Amplification trials differed by annealing temperature, amount of DNA and primer used, and sample type. Results of amplification were verified through gel electrophoresis and visible bands of approximately size 83-99 bp. The results on CSRM60 efficacy contribute to a baseline for future researchers to initiate a population-level genetic analysis of the remaining anoa that will provide identification of individuals and assess genetic diversity, ultimately informing conservation strategies for the endangered anoa.
Amplifying the Anoa: Noninvasive Genetic Amplification of the Lowland Anoa with CSRM60
The lowland anoa (Bubalus depressicornis) is the smallest species of wild cattle in the world and is classified as endangered on the ICUN Red List. Noninvasive genetic assessment allows for collection of samples with minimal impact on animal welfare but can result in low-quality DNA or contamination. Little is known about effective forms of noninvasive sampling and the DNA analysis protocol that follows regarding the anoa. This study is part of a continuing collaborative investigation and helped gain insight into the suitability of noninvasive DNA for population research. To determine whether the DNA is suitable, a set of genetic markers is needed. This study tested whether noninvasive DNA from fecal and saliva samples would amplify using primer CSRM60. CSRM60 was selected based on cross-species amplification in bovids, (e.g., cattle and domestic buffaloes). CSRM60 targets a microsatellite locus with an allele size range of 83-99 bp. This study primarily tested DNA taken from fecal samples stored in ethanol and extracted using a Qiagen Fast DNA Kit. Saliva samples tested were stored in DNA/RNA shield and extracted by Quick-DNA MICROprep Kit. Amplification trials differed by annealing temperature, amount of DNA and primer used, and sample type. Results of amplification were verified through gel electrophoresis and visible bands of approximately size 83-99 bp. The results on CSRM60 efficacy contribute to a baseline for future researchers to initiate a population-level genetic analysis of the remaining anoa that will provide identification of individuals and assess genetic diversity, ultimately informing conservation strategies for the endangered anoa.
