BME100 f2013:W900 Group14 L4: Difference between revisions
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http://openwetware.org/images/thumb/e/ef/HALEY.png/562px-HALEY.png<br> | http://openwetware.org/images/thumb/e/ef/HALEY.png/562px-HALEY.png<br> | ||
http://www.wired.com/reviews/wp-content/uploads/2011/12/reviews_pcr_f.jpg<br> | http://www.wired.com/reviews/wp-content/uploads/2011/12/reviews_pcr_f.jpg<br> | ||
PCR stands for polymerase chain reaction. The PCR machine depicted here is a biochemical technology that when used properly can amplify a selected piece of | PCR stands for polymerase chain reaction. The PCR machine depicted here is a biochemical technology that when used properly can amplify a selected piece of DNA to make millions of copies. This machine heats DNA to break it apart, then cools it so that the polymerase will attach to the DNA strands and make copies. For the heating process, the Open PCR machine is built with highly thermally conductive alloy of aluminum and insulation, allowing heat to be kept in the machine as well as spread the temperature across the block and be able to affect the test tubes. With a adjustable lid on top of the machine, condensation of water in the test tubes is essentially negated, allowing correct and reproducible results to appear. Last but not least, the Open PCR machine has the ability to connect to essentially any computer, compatible with a Windows or Mac though USB. Also the GUI program is only necessary for the initial and final steps, allowing a disconnection from the PC. The LED machine indicates the status of reaction, such as temperature, cycles, as well as remaining time until the reaction is complete. Finally, due to a file system interface via USB, the Open PCR machine can integrate into lab or other external control systems. | ||
'''Experimenting With the Connections'''<br> | '''Experimenting With the Connections'''<br> | ||
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{| {{table}} | {| {{table}} | ||
|- | |- | ||
| | | Cancer DNA Template TUBE#: CON+|| Positive Patient ID: 86763 TUBE#:1a || ID:86763 TUBE#:1b || ID:86763 TUBE#:1e | ||
|- | |- | ||
| | | NON-Cancer DNA Template TUBE#: CON-|| Negative Patient ID: 65583 TUBE#:2a|| ID:65583 TUBE#:2b || ID:65583 TUBE#:2e | ||
|} | |} | ||
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'''DNA/ primer mix''' | '''DNA/ primer mix''' | ||
* | * Within the DNA/Primer mix is the sample of DNA with the desired strands to be replicated as well as RNA primer. | ||
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'''PCR - The Underlying Technology'''<br> | '''PCR - The Underlying Technology'''<br> | ||
THE COMPONENTS OF PCR<br> | |||
There are three steps in PCR, these steps are: 1)Denaturation, 2)Annealing, and 3)Extension. | |||
During the Denaturation, the DNA samples are heated to slightly under the boiling point of water, or around 94 degrees C. At this point the DNA strands break apart at the hydrogen bonds between the complementary bases, into the two basic strands that compose non-denatured DNA. During the annealing step, the DNA sample is cooled to around 54 degrees C where the forward and reverse primers attach on the single stranded DNA as the polymerase binds them, so as to allow the DNA Taq Polymerase to bind to the strands and create complementary copies. Finally, during the extension phase, the DNA is reheated to 72 degrees Farenheit, so as to stimulate the polymerase base pair copying reaction to build new DNA. | |||
Latest revision as of 08:59, 30 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 (part 3) from (part 6), the machine's screen went black. The LCD screen did not receive power while the rest of the machine still did. When we unplugged the white wire that connects (part 6) to (part 2), the machine did not read temperature properly. It briefly fluctuating upon detaching the wire, then read -40 degrees C.
Wednesday, October 23, 2013
ProtocolsThermal Cycler Program
PCR Reaction Mix
DNA/ primer mix
Research and DevelopmentPCR - The Underlying Technology THE COMPONENTS OF PCR
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