BME103 s2013:T900 Group4: Difference between revisions
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==Research and Development== | ==Research and Development== | ||
Mixture components: | |||
DNA Template <br> | DNA Template (Sample DNA)<br> | ||
DNA Polymerase<br> | DNA Polymerase<br> | ||
DNA Primers<br> | DNA Primers<br> | ||
Nucleotides<br> | Free Nucleotides<br> | ||
Procedure:<br> | Procedure:<br> | ||
First, the | First the DNA sample, polymerase, primers, and nucleotides are combined in a small test tube. Repeat samples are advised to make sure that the results are consistent. When the test tube is placed in a PCR machine, the mixture is heated to 95 degrees Celsius for 30 seconds. This high heat breaks apart complementary strands of DNA, exposing the nucleotides. The machine then rapidly cools to 55 degrees Celsius for 30 seconds, allowing the DNA primers to bind to the exposed sites. The DNA primers can only bind to the specific site they are complementary to, which allows researchers to target specific DNA sequences as discussed below in the cancer detection section. The mixture is then heated up to 82 degrees Celsius for 30 seconds. At this temperature, DNA polymerase binds to the primers and builds a new segment of DNA with the free nucleotides in the mixture. This new strand of DNA is complementary to the original. So, if there was only one strand of DNA in the mixture originally, now there are two (since both halves have been replicated). The mixture is then reheated to 95 degrees and the cycle continues, each time doubling the amount of DNA in the mixture. This process usually continues for an hour or so, until there are billions of copies of DNA. This whole process is referred to as DNA amplification. | ||
'''Specific Cancer Marker Detection - The Underlying Technology'''<br> | '''Specific Cancer Marker Detection - The Underlying Technology'''<br> |
Revision as of 23:44, 25 March 2013
BME 103 Spring 2013 | 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 ... (did what? fill in your answer) When we unplugged the white wire that connects (part 6) to (part 2), the machine ... (did what? fill in your answer)
(Write the date you first tested Open PCR and your experience(s) with the machine)
ProtocolsThermal Cycler Program DNA Sample, 50 μL each: patient ID's from the UnderGrad Teaching Assistant(UGTA).
DNA Sample Set-up Procedure
DNA/ primer mix
Research and DevelopmentMixture components: DNA Template (Sample DNA) Procedure: Specific 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.)
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