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

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

3. Hypothesis TT he 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

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 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 DNA Extraction Mouse Tails Qiagen DNeasy Protocol Thermal Cycler (PCR Machine)

8. PCR Procedure

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

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_g enotyping.asp  http://en.wikipedia.org/wiki/Mouse  http://en.wikipedia.org/wiki/Genotyping  http://www.skincancer.org/melanoma/index.php

12. Acknowledgements  Dr. Taha Merghoub  Diana  Hong  Houghton Laboratory  Dr. Sat Bhattacharya  Harlem Children Society