BME103:T930 Group 11: Difference between revisions
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'''Polymerase Chain Reaction'''<br> | '''Polymerase Chain Reaction'''<br> | ||
The Polymerase Chain Reaction works by attaching MgCl to taq enzyme in the solution and then pulling deoxy nucleotide diphosphates, binding them to the strand that is already present. This gives a base to the DNA that is already there.<br>DNA amplification contains three important steps: heat denauturation,primer annealing, and primer extension. <br> Step One: Heat Denauturation-Heat is applied to the DNA and the two strands separate. <br>Step Two:Primer Annealing-With excess dNTPs, oligonucleotides are added.<br>Step three: Primer Extension-DNA polymerase is added and new strands of DNA are synthesized. The DNA strands combine with the nucleotides to form completed, complimentary strands of DNA. <br>PCR master mix contains Magnesium Chloride, all four Nucleotides needed to create DNA, and reaction buffers.<br>Reagent volume<br>Template DNA(20 ng) .2''u''L<br>10 ''u''M forward primer 1.0 uL<br>10 uM reverse primer 1.0 uL<br>GoTaq master mix 50.0 uL<br>DH2O 47.8 uL<br> Total Volume 100.0 uL<br> | The Polymerase Chain Reaction works by attaching MgCl to taq enzyme in the solution and then pulling deoxy nucleotide diphosphates, binding them to the strand that is already present. This gives a base to the DNA that is already there.<br>DNA amplification contains three important steps: heat denauturation,primer annealing, and primer extension. <br> Step One: Heat Denauturation-Heat is applied to the DNA and the two strands separate. <br>Step Two:Primer Annealing-With excess dNTPs, oligonucleotides are added.<br>Step three: Primer Extension-DNA polymerase is added and new strands of DNA are synthesized. The DNA strands combine with the nucleotides to form completed, complimentary strands of DNA. <br>PCR master mix contains Magnesium Chloride, all four Nucleotides needed to create DNA, and reaction buffers.<br><br.<b>Chart One: Reagents and Their Volumes</b><br>Reagent volume<br>Template DNA(20 ng) .2''u''L<br>10 ''u''M forward primer 1.0 uL<br>10 uM reverse primer 1.0 uL<br>GoTaq master mix 50.0 uL<br>DH2O 47.8 uL<br> Total Volume 100.0 uL<br> | ||
<br>Patient one's ID number is 12123, male, and 61 years old. Patient two's ID is 21312, female, and 56 years old. | <br>Patient one's ID number is 12123, male, and 61 years old. Patient two's ID is 21312, female, and 56 years old. | ||
Revision as of 22:44, 14 November 2012
BME 103 Fall 2012 | Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help | ||||||||||||||||||||||||||||||||||||||||||
OUR TEAMLAB 1 WRITE-UPInitial Machine TestingThe Original Design This is an Open PCR machine. The purpose of the machines is to fluctuate the temperature of different samples of DNA strands to separate them into a single strand. It allows for a way to amplify DNA sequences by this separation and heating them with primers and DNA polymerase. When raised to a high enough temperature DNA melts, which is where it separates into the two strands, and primers and DNA polymerase are used to fill in the holes and attach to the now single-stranded DNA. The machine is able to amplify a specific sequence of DNA up to 1 billion times. Because of this ability, scientists and investigators can use it to amplify these sequences even if only a small amount of DNA is provided. Single roots in hair or even microscopic splatters of blood left such as at scenes of crime are actually ample amounts of DNA for PCR.
When we unplugged (part 3) from (part 6), the machine's display screen didn't turn on or show up. When we unplugged the white wire that connects (part 6) to (part 2), the machine's temperature reading was altered compared to the accurate values from the computer.
ProtocolsPolymerase Chain Reaction The Polymerase Chain Reaction works by attaching MgCl to taq enzyme in the solution and then pulling deoxy nucleotide diphosphates, binding them to the strand that is already present. This gives a base to the DNA that is already there.
Flourimeter Measurements
Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology (Add a write-up of the information discussed in Week 3's class) (BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)
Results
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