1. Evaluating the Genetic Health in Bison on the Pine Ridge Reservation, SD
Tada R Vargas
Oglala Lakota College
Mentor: Alessandra Higa
NIFA-USDA award #

2. Overview Background Target Population Research Question Methodology
Results
Conclusion

3. Cultural Significance

4. History

5. Diagram 1: Founder’s Effect
Restoration Process
Today, North American bison populations exhibit a range of genetic purity depending on the history and management of individual herds. Determining the purity of a population is vital because, at some point, the level of introgression will impact the behavior, reproductive viability, and overall genetic species of an individual.
Today, North American bison populations exhibit a range of genetic purity depending on the history and management of individual herds.
Determining the purity of a population is vital because, at some point, the level of introgression will impact
the behavior,
reproductive viability, and
overall genetic species
Graph 1: Bottle Necking
Diagram 1: Founder’s Effect

6. Research Question: Hypothesis: Null Hypothesis:
How has cattle introgression affected the genetic diversity in Bison on the Pine Ridge Reservation?
Hypothesis:
The genetic variation of this population will not be in equilibrium from generation to generation due to disturbing factors.
Null Hypothesis:
The genetic variation of this population will remain constant from generation to generation.

7. Oglala Sioux Parks and Recreation Association

8. Methodology Hair Follicle DNA Extraction Mitochondrial based PCR assay
Nuclear based PCR assay
Electrophoresis
Genotyping
Genetic Analysis
GenePop: Hardy Weineberg, Relatedness, and Inbreeding Coefficient

9. Figure 2: Extracting DNA
DNA Extraction
What happens to the cell. Cell breaks open, lipids, proteins, and RNA removed and DNA is purified.
Protease, RNase, TE Buffer
Ethanol and NaOH
Figure 2: Extracting DNA
Figure 3: Extracted DNA

10. Polymerase Chain Reaction PCR
PCRReagents: dNTPs, BSA, Taq, MgCl2, and water
Figure 4: Aliquoting Master Mix
Figure 5: Transferring DNA
Figure 6: Thermocycler
Figure 7: Heating and Cooling Process

11. Figure 9: Gel Electrophoresis
Gel electrophoresis is a method used to separate and analyze DNA fragments, based on their size and charge.
They are separated by applying an electric field, which moves the negatively charged molecules thru the gel.
Of course, smaller molecules will move faster than larger ones and this is how it is separated by size.
After we view the gel in the photodyne to verify a successful reaction.
What time of light is used in photodyne?
Figure 9: Gel Electrophoresis
Figure 10: Photodyne

12. Maternally Inherited Introgression
Figure 11: Controls
Figure 12: Gel results for Mitochondria analysis

13. Table 1 Nuclear Primers of PCR Multiplex Analysis
Paternally Inherited Introgression
Table 1 Nuclear Primers of PCR Multiplex Analysis
Figure 13 and Figure 14: Gel photo for Nuclear Panel Primers

14. Figure 16 and 17: GeneAnalyzer (Closed and Open)
Genotyping
Prepping samples for genotyping is much like PCR in that we subject samples to reagents only in PCR we are amplifying a desired fragment.
In genotyping, using the Geneanalyzer, we are able to discern what that fragment looks like.
In this case are they homozygous bison, heterozygous bison, or homo or heterozygous cattle
Figure 15: Aliquoting dilute PCR product for genotyping
Figure 16 and 17: GeneAnalyzer (Closed and Open)

15. Introgression Genotypes Plot 1: No Introgression Confirming Markers
Primer: RM 185
*Bison at 92 bp
*Cattle between
Genotypes Plot 2:
Positive Cattle Introgression
Primer: BM 4307
*Bison between
*Cattle between

17. Variant Alleles Genotype Plots 5: Primer CSSM42
Bison: 167 – 171 Cattle:
Genotype Plots 6:
Primer BM4307
Bison: 185 – 187 Cattle:
Genotype Plots 7:
Primer CSSM42
Bison: 167 – 171 Cattle:

18. Genetic Diversity Genotype Plot 8: Confirming Marker Primer RM500
Bp size Bison: 123 Cattle

19. Genetic Diversity GenePlot: Confirming Marker INRA 119
Bp size Bison: Cattle:

20. Genetic Diversity Genotype Plot 9: Introgression Primer BM4307
Bison: 185 – 187 Cattle:

21. Conclusion
Genotyping is necessary to determine the presence of introgression.
Exact levels of introgression cannot be determined without genotyping the entire genome.
Possible new alleles but cannot be determined without genetic sequencing
Provide detailed knowledge to develop strategies to best suit management goals
Beneficial to interspecies introgression for wildlife population management and species conservation.

22. Acknowledgements

23. Questions?