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Genotyping Transgenic Mice

Genotyping Transgenic Mice. By: Natalie Saldana & Dr. Taha Merghoub. Introduction. Human Disease. Melanoma - serious form of skin cancer. Mouse Model – used to mimic the human disease. Transgenic Mouse. Hypothesis.

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Genotyping Transgenic Mice

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  1. Genotyping Transgenic Mice By: Natalie Saldana & Dr. Taha Merghoub

  2. Introduction Human Disease Melanoma - serious form of skin cancer MouseModel – used to mimic the human disease Transgenic Mouse

  3. Hypothesis • The introduction of Human melanoma mutation in mice will induce this disease in mice

  4. Why Mice? • They are genetically similar to human beings • Affordable and easy to maintain • Able to reproduce within 3 weeks • Short life span • Small

  5. Transgenic Mouse Genotype

  6. Genotyping • The process of testing individuals in order to reveal any specific gene that has been inherited from that individual • Includes other methods like PCR, DNA sequencing, and Southern blot • The goal is to understand the functions and roles of genes and to test new therapies in melanoma

  7. Materials DNA Extraction Mouse Tails Qiagen DNeasy Protocol PCR Taq Polymerase Buffer MgCl2 Primers dNTPs DNA H2O Gel Electrophoresis Agarose TAE Buffer Ethidium Bromide Trays & Combs Gel Box Power Supply UV Light 1 Kb Ladder Loading Dye Thermal Cycler (PCR Machine)

  8. PCR Procedure

  9. Observations & Results  Ink4A WT We were successful in genotyping the mice which we will use for future experiments.  Ink4A KO

  10. Conclusion and Future Directions • We have identified Heterozygous and KO Ink4a mice. • The Ink4a mice will be crossed with NRas transgenic mice in order to obtain a mouse with melanoma.

  11. Reference • http://users.ugent.be/~avierstr/principles/pcr.html • http://www.accessexcellence.org/RC/VL/GG/transgenic.html • http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/T/TransgenicAnimals.html • http://sunsite.berkeley.edu/PCR/whatisPCR.html • http://www.agrf.org.au/Default.aspx?tabid=85 • http://ghr.nlm.nih.gov/ghr/glossary/genotyping • http://www.identigene.com/SWIMX/Products/mouse_genotyping.asp • http://en.wikipedia.org/wiki/Mouse • http://en.wikipedia.org/wiki/Genotyping • http://www.skincancer.org/melanoma/index.php

  12. Acknowledgements THANK YOU! • Dr. Taha Merghoub • Diana • Hong • Houghton Laboratory • Dr. Sat Bhattacharya • Harlem Children Society

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