Genotyping transgenic mice
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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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Genotyping transgenic mice

Genotyping Transgenic Mice

By: Natalie Saldana

&

Dr. Taha Merghoub


Introduction

Introduction

Human Disease

Melanoma - serious form of skin cancer

MouseModel – used to mimic the human disease

Transgenic Mouse


Hypothesis

Hypothesis

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


Why mice

Why Mice?

  • They are genetically similar to human beings

  • Affordable and easy to maintain

  • Able to reproduce within 3 weeks

  • Short life span

  • Small


Transgenic mouse

Transgenic Mouse

Genotype


Genotyping

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


Materials

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)


Pcr procedure

PCR Procedure


Observations results

Observations & Results

 Ink4A WT

We were successful in genotyping the mice which we will use for future experiments.

 Ink4A KO


Conclusion and future directions

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.


Reference

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


Acknowledgements

Acknowledgements

THANK YOU!

  • Dr. Taha Merghoub

  • Diana

  • Hong

  • Houghton Laboratory

  • Dr. Sat Bhattacharya

  • Harlem Children Society


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