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> | 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>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 19:18, 13 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
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.
(Write the date you first tested Open PCR and your experience(s) with the machine)
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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