BME100 f2013:W900 Group13 L4: Difference between revisions
Line 27: | Line 27: | ||
'''The Original Design'''<br> | '''The Original Design'''<br> | ||
[[Image: | [[Image:allisonopenpcr.jpg]] | ||
The OpenPCR machine runs PCR reactions and can be monitored by a computer to which the machine can be connected with a USB 2.0 cable. The machine has a heated lid which prevents condensation on the caps of the PCR tubes. If this part did not work, water would condensate on the cap and cause the salt concentration to be too high for the polymerase to function properly. The heated lid is situated over the 16 tube PCR block, which holds and heats the samples in the PCR tubes. If the block did not work, the samples would not be heated to the necessary temperatures and there would not be a PCR reaction. Information about the reaction is displayed on the screen located on top of the box. Inside the box there are the heating components for the PCR block. These heating components go to a heat sink which leads into an air filter which a fan moves air through so as to keep the temperature within the box from getting too hot and the unit from overheating. The wiring for the PCR block, the cooling system, and the display are all hooked to a motherboard situated at the bottom of the box. The motherboard controls the temperature through a connection to the PCR block and runs the OpenPCR machine. <br> | The OpenPCR machine runs PCR reactions and can be monitored by a computer to which the machine can be connected with a USB 2.0 cable. The machine has a heated lid which prevents condensation on the caps of the PCR tubes. If this part did not work, water would condensate on the cap and cause the salt concentration to be too high for the polymerase to function properly. The heated lid is situated over the 16 tube PCR block, which holds and heats the samples in the PCR tubes. If the block did not work, the samples would not be heated to the necessary temperatures and there would not be a PCR reaction. Information about the reaction is displayed on the screen located on top of the box. Inside the box there are the heating components for the PCR block. These heating components go to a heat sink which leads into an air filter which a fan moves air through so as to keep the temperature within the box from getting too hot and the unit from overheating. The wiring for the PCR block, the cooling system, and the display are all hooked to a motherboard situated at the bottom of the box. The motherboard controls the temperature through a connection to the PCR block and runs the OpenPCR machine. <br> | ||
Revision as of 15:58, 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
When we unplugged (part 3) from either end, the machine's screen turned off. When we plugged the cord back in, the screen turned back on. We believe this part supplies power to the screen. When we unplugged the white wire that connects (part 6) to (part 2), the machine showed no noticeable change. However, we believe that the temperature controls for the thermocycler will not work when the white wire is disconnected. Test Run
Test Date: 10/23/13
Experience: The Open PCR's time functioned properly, but the component that keeps track of the cycles did not. The unit indicated that it was on the first cycle of thirty-five throughout the duration of the PCR reaction.
(Write the date you first tested Open PCR and your experience(s) with the machine)
ProtocolsThermal Cycler Program 1. Begins with the initial denaturation step which consists of one cycle at 95C for three minutes 2. Moves next to the denaturation step which consists of 35 cycles at 95C for 30 seconds, the anneal step which includes 57C for 30 seconds, and the extended step including 72C for 30 seconds for each cycle. 3. The Final extension step occurs at 72C for 3 minutes 4. The refrigeration of the DNA occurs during the final hold at 4C
DNA Sample Set-up Procedure 1. Step 1: Label each tube with its respective contents to prevent misdiagnosing patients. (Refer to Table #1 for labelling)
PCR Reaction Mix •Taw DNA Polymerase •MgCl2 •dNTP
•A Forward Primer •A Reverse Primer Six of the eight tubes will contain unique DNA samples from two different patients. The remaining two tubes will contain control templates with and without cancerous DNA: the Positive Control Tube will contain the cancerous DNA and the Negative will not.
Research and DevelopmentPCR - The Underlying Technology (Add a write-up, essay-style, organized into paragrpahs with descriptive headers, based on the Q&A's from Section three of your worksheet) (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.) What is the function of each component of a PCR reaction?
|