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Fluorescence to Study Genetic Material

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Roslyn Mainland

on 4 February 2014

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Transcript of Fluorescence to Study Genetic Material

Fluorescence to Study Genetic Material
How do objects fluoresce?
How do specimens fluoresce?
Advantages of Fluorescence Microscopy
high contrast
fluorochromes
live imaging
3D structure
Fluorescent Microscopy Applications
1. Excitation Light
2. Emission Light
Excitation vs Emission Light
How does a fluorescent microscope work?
Emission of Light
fluorochromes (stains)
1. DNA
Identification
2. DNA Sequencing
photobleaching
Disadvantages of
Fluorescence Microscopy
to create map of human genome
chain termination method and shotgun technique
available to researchers to locate and research specific genes
Article 1: Revealing the competition between peeled ssDNA, melting bubbles, and S-DNA during DNA overstretching using fluorescence microscopy

DNA overstretching = lengthening due to mechanical stress
Three Mechanisms:
strand unpeeling
melting bubble formation
S-DNA formation
Three Factors:
DNA sugar-phosphate backbone structure
DNA sequence
ionic strength
Article 2: Human Amnion Epithelial Cells Induced to Express Functional Cystic Fibrosis Transmembrane Conductance Regulator
Cystic Fibrosis =
mutation of CFTR gene
Cellular Therapy =
injection of material, like epithelial cells, into patient to treat disease

Cells demonstrated CFTR gene:
protein synthesis
3D structure
functionality
Technical Aspect
Application
Article Summaries
intercalation
groove (external)
binding
fluorescent
probes
3. Karyotyping
DNA Identification:
- achieved through Gel Electrophoresis
Results from a gel electrophoresis test:
Gel electrophoresis technique:
= DNA fragments are separated to determine size
forensics, paternity/maternity tests, DNA sequencing
DNA Sequencing:
= determining base sequences of DNA
Method 1:
Chain Termination Method
Method 2:
Shotgun Technique
DNA cut into tiny fragments
cloning
computer software
Human Genome Project
Predict Gene Function
comparison of unknown gene to similar gene with known function
Determine Evolutionary History
comparison of similar genes between species
Karyotyping:
= map of person's 23 pairs of chromosomes
Procedure:
Karyotype Example
Identifies:
deletions
duplications
inversions
translocations
crossing-over
nondisjunction
What is fluorescence
microscopy?
diagnoses genetic disorders
Bibliography
Rice, George. “Fluorescent Microscopy.” Microbial Life. Science Education Resources Center,
19 Nov. 2013. Web. 27 Jan. 2014
<http://serc.carleton.edu/microbelife/research_methods/microscopy/fluromic.html>.

Abramowitz, Mortimer, and Michael W. Davidson. “Introduction to Fluorescence.” Olympus
Microscopy Resource Center. Olympus America Inc., n.d. Web. 27 Jan. 2014
<http://www.olympusmicro.com/primer/lightandcolor/fluorointroduction.html>.

“Fish Fact Sheet”. genome.gov. National Human Genome Research Institute, 13 Oct. 2011.
Web. 27 Jan. 2014 <http://www.genome.gov/10000206>.

Spector, David L., Robert D. Goldman, and Leslie A. Leinwand. Cells: a laboratory manual.
Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, 1998. Print.

“An Overview of the Human Genome Project.” genome.gov. National Human Genome Research
Institute, 8 Nov. 2012. Web. 25 Jan. 2014 <http://www.genome.gov/12011238>.

O’Connor, Clare. “Karyotyping for Chromosomal Abnormalities.” Scitable. Nature Publishing
Group, n.d. Web. 27 Jan. 2014 <http://www.nature.com/scitable/topicpage/karyotyping-
for-chromosomal-abnormalities-298>.

Alberts, Bruce. Molecular biology of the cell. 4th ed. New York: Garland Science, 2002. Print.

“Using Karyotypes To Diagnose Genetic Disorders.” Learn.Genetics. University of Utah, n.d.
Web. 27 Jan. 2014. <http://learn.genetics.utah.edu/content/chromosomes/diagnose>.

Davidson, Michael W. “The Future of Fluorescence Microscopy.” Zeiss. Carl Zeiss, n.d. Web.
27 Jan. 2014.
<http://www.zeiss.com/C125716F004E0776/0/87618A6A573E028FC125755A0047E65
C/$File/inno-en_21_30long.pdf>.

Edelson, Edward. Genetics and heredity. New York: Chelsea House, 1990. Print.
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