genotyping a transgenic mouse ink4a knock-out & ink4a wild-type
DESCRIPTION
Genotyping A Transgenic Mouse INK4A Knock-out & INK4A Wild-type. 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. Melanoma. - PowerPoint PPT PresentationTRANSCRIPT
Genotyping A Transgenic MouseINK4A Knock-out & INK4A Wild-type
By: Natalie Saldana &
Dr. Taha Merghoub
Introduction
Mouse Model – used to mimic the human disease
Melanoma - serious form of skin cancer
Transgenic Mouse
Human Disease
Melanoma A rare type of skin cancer that originates in the
melanocytes - found predominantly in skin but also in the bowel and the eye
Causes: Sun exposure - UVA and UVB rays are can
induce skin cancer Moles - normal moles — the small brown
blemishes, growths, or "beauty marks" atypical moles -- also known as dysplastic nevi
Skin Type - fairer skin are at increased risk Family History Personal History - Once you have had melanoma,
you run an increased chance of recurrence Weakened Immune System - immune systems
effected with chemotherapy, an organ transplant, excessive sun exposure, and diseases such as HIV/AIDS or lymphoma can increase your risk of melanoma
Why Mice?
They are genetically similar to human beingsAffordable and easy to maintainAble to reproduce within 3 weeksShort life spanSmall
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 PCR
Taq PolymeraseBufferMgCl2PrimersdNTPsDNAH2O
Gel ElectrophoresisAgarose
TAE BufferEthidium Bromide
Trays & CombsGel Box
Power SupplyUV Light
1 Kb LadderLoading Dye
DNA Extraction
Mouse Tails
Qiagen DNeasy Protocol
Thermal Cycler (PCR Machine)
Observations & Results
Ink4A WT
Ink4A KO
We were successful in genotyping the mice which we will use for future experiments.
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
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