3. What is in the master mix and why do you need each component? 4. Why do you need to perform PCR on DNA evidence from a crime scene? 5. What steps make up a PCR cycle, and what happens at each step?

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Chapter17: Biotechnology And Genomics
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Problem 5RQ: Gene therapy can be used to introduce foreign DNA into cells. for molecular cloning by PCR of...
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IMAGINE THE FOLLOWING SCENARIO
Scene: The Highway Motel, #1 Dark Highway, Nowhere Setting: Room #13.
The motel manager hears loud voices, a woman screams, and a shot rings out. The manager runs to
the window in time to see the receding lights of a car leaving in a hurry. The door to room #13 hangs
open. The manager runs to the open door to see a man lying face down in a pool of blood. He calls
911. The police arrive and begin to examine the crime scene. An apparent homicide, but with no
obvious clues as to who committed the crime.
Or...?
A forensic specialist is called in to examine the crime scene and collect evidence. Even though it looks like
the people involved left no evidence behind, the specialist can use laboratory tests that can tell who was
at the crime scene from a single drop of blood or a lone hair. Is this a science fiction story, or reality?
Very much a reality. Testing is routinely done in forensic testing labs across the U.S. and in many other
parts of the world from only a single cell, and sometimes from samples that are decades old. The
reason this is possible is because of DNA. To be able to perform laboratory tests, the specialist needs
biological material to work with. Often, there is very little material left at the scene of a crime, and not
in quantities that will allow analysis. To get around this problem, the specialist takes advantage of a
process that each and every cell in your body uses to divide.
The most important part of any cell's life is when it commits to reproducing itself and dividing. The
basic result of any cell division is the creation of two identical daughter cells from one original cell. To
ensure that this happens, DNA replication must have a high degree of specificity and accuracy, that
is, it must copy DNA exactly. To do so, the enzymes involved in DNA replication use the information
already contained in the existing strands to make new DNA copies. This basic idea - the exact
copying of DNA from a template - is the basis for a new technology that has revolutionized many
areas of science, medicine, and the courts.
PCR allows the forensic specialist to specifically amplify, or copy, any region of DNA that he or she is
interested in. PCR is the basis for DNA testing that is currently used in nearly all forensic analysis.
In this experiment, you will perform PCR analysis on a single locus, the BXP007 locus, using template
DNAS obtained from a simulated crime scene and a victim. Following PCR, you will run an agarose
gel to separate the PCR products, visualize the PCR products, compare them to a simulated ladder of
possible alleles for this locus, and assign a genotype for the templates. You will then look to see if any
of the suspects' genotype match the crime scene, and see whether you can determine whodunit!
Let's examine the DNA evidence and find out who pulled the trigger.
172
Introduction to PCR- Based Forensics
Transcribed Image Text:IMAGINE THE FOLLOWING SCENARIO Scene: The Highway Motel, #1 Dark Highway, Nowhere Setting: Room #13. The motel manager hears loud voices, a woman screams, and a shot rings out. The manager runs to the window in time to see the receding lights of a car leaving in a hurry. The door to room #13 hangs open. The manager runs to the open door to see a man lying face down in a pool of blood. He calls 911. The police arrive and begin to examine the crime scene. An apparent homicide, but with no obvious clues as to who committed the crime. Or...? A forensic specialist is called in to examine the crime scene and collect evidence. Even though it looks like the people involved left no evidence behind, the specialist can use laboratory tests that can tell who was at the crime scene from a single drop of blood or a lone hair. Is this a science fiction story, or reality? Very much a reality. Testing is routinely done in forensic testing labs across the U.S. and in many other parts of the world from only a single cell, and sometimes from samples that are decades old. The reason this is possible is because of DNA. To be able to perform laboratory tests, the specialist needs biological material to work with. Often, there is very little material left at the scene of a crime, and not in quantities that will allow analysis. To get around this problem, the specialist takes advantage of a process that each and every cell in your body uses to divide. The most important part of any cell's life is when it commits to reproducing itself and dividing. The basic result of any cell division is the creation of two identical daughter cells from one original cell. To ensure that this happens, DNA replication must have a high degree of specificity and accuracy, that is, it must copy DNA exactly. To do so, the enzymes involved in DNA replication use the information already contained in the existing strands to make new DNA copies. This basic idea - the exact copying of DNA from a template - is the basis for a new technology that has revolutionized many areas of science, medicine, and the courts. PCR allows the forensic specialist to specifically amplify, or copy, any region of DNA that he or she is interested in. PCR is the basis for DNA testing that is currently used in nearly all forensic analysis. In this experiment, you will perform PCR analysis on a single locus, the BXP007 locus, using template DNAS obtained from a simulated crime scene and a victim. Following PCR, you will run an agarose gel to separate the PCR products, visualize the PCR products, compare them to a simulated ladder of possible alleles for this locus, and assign a genotype for the templates. You will then look to see if any of the suspects' genotype match the crime scene, and see whether you can determine whodunit! Let's examine the DNA evidence and find out who pulled the trigger. 172 Introduction to PCR- Based Forensics
3. What is in the master mix and why do you need each component?
4. Why do you need to perform PCR on DNA evidence from a crime scene?
5. What steps make up a PCR cycle, and what happens at each step?
176 Introduction to PCR-Based Forensics
Transcribed Image Text:3. What is in the master mix and why do you need each component? 4. Why do you need to perform PCR on DNA evidence from a crime scene? 5. What steps make up a PCR cycle, and what happens at each step? 176 Introduction to PCR-Based Forensics
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