BME100 f2013:W1200 Group13 L4: Difference between revisions
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A Polymerize Chain Reaction is a way of making various copies of DNA molecules. A PCR reaction has 5 different components and each of these components have a different function. The first component is the template DNA, this is the sample that contains the sequence that we want to replicate. Then there's the primers,short pieces of synthesized DNA, that bond to the target sequence and are the starting point for replication. The Taq Polymerase physically constructs the new strand of DNA. In the reaction, Magnesium Chloride facilitates the bonding of the nucleotides and the Deoxyribonucleotides are converted to dNMP's by the magnesium chloride before bonding to another nucleotide. <br> | A Polymerize Chain Reaction is a way of making various copies of DNA molecules. A PCR reaction has 5 different components and each of these components have a different function. The first component is the template DNA, this is the sample that contains the sequence that we want to replicate. Then there's the primers,short pieces of synthesized DNA, that bond to the target sequence and are the starting point for replication. The Taq Polymerase physically constructs the new strand of DNA. In the reaction, Magnesium Chloride facilitates the bonding of the nucleotides and the Deoxyribonucleotides are converted to dNMP's by the magnesium chloride before bonding to another nucleotide. <br> | ||
At the beginning of the thermal cycling, the samples are heated for 3 minutes at a temperature of 95 degree Celsius | At the beginning of the thermal cycling, the samples are heated for 3 minutes at a temperature of 95 degree Celsius, the heat makes the double-helix unwind and separate into two single stranded DNA molecules. Afterwards, the temperature is brought down to 57 degrees Celsius, the temperature change makes the target attach to the target sequence. Once again, the temperature is brought up, this time to 72 degrees celsius, this activated the TAQ polymerase and attaches to the primer that is attached to the single DNA strand and adds complimentary nucleotides to the strand until it detaches. Now, we have a copy of the specific sample we wanted, in this case the cancer mutations. Doing this helps because it only gives the cancer mutation and scientists can work and figure out what is wrong with that piece of DNA and try to solve the problem. | ||
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Revision as of 20:37, 29 October 2013
BME 100 Fall 2013 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | ||||||||||||||
OUR TEAMLAB 1 WRITE-UPInitial Machine TestingThe Original Design Experimenting With the Connections When we unplugged the LED screen (part 3) from the circuit board(part 6), the machine didn't display anything on the screen because this connection is what allows the power to connect to the led screen. When we unplugged the white wire that connects (part 6) to (part 2), the machine lost the ability to record data and to sense the heat change in the machine.
Our OpenPCR machine passed the testing that we conducted in class. We began the testing at 12:50 pm, and at 1:40pm the machine had run 19 out of 35 tests. When we concluded the experiment the temperature according to the screen on the device was 95 degrees Celsius, and had an ETA of 48. The test ran very smoothly for the first time use and we didn't encounter problems with the machine not working properly. The device was plugged into the computer and then we opened the program in order to start the tests. Then after entering in the information given we began running the test. There were no issues with the program or the machine. )
ProtocolsThermal Cycler Program DNA Sample Set-up
DNA Sample Set-up Procedure
The PCR reaction , 50uL each. the mix contains Taq DNA polymerase, MGCl2, and dNTP's. DNA/ primer mix
Each mix contains a differant template DNA all tubes have the same forward primer and reverse primer.
Research and DevelopmentPCR - The Underlying Technology At the beginning of the thermal cycling, the samples are heated for 3 minutes at a temperature of 95 degree Celsius, the heat makes the double-helix unwind and separate into two single stranded DNA molecules. Afterwards, the temperature is brought down to 57 degrees Celsius, the temperature change makes the target attach to the target sequence. Once again, the temperature is brought up, this time to 72 degrees celsius, this activated the TAQ polymerase and attaches to the primer that is attached to the single DNA strand and adds complimentary nucleotides to the strand until it detaches. Now, we have a copy of the specific sample we wanted, in this case the cancer mutations. Doing this helps because it only gives the cancer mutation and scientists can work and figure out what is wrong with that piece of DNA and try to solve the problem.
(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 PCR video/ tutorial might be useful. Be sure to credit the sources if you borrow images.)
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