BME100 f2013:W900 Group13 L4: Difference between revisions
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1. Step 1: Label each tube with its respective contents to prevent misdiagnosing patients. (Refer to Figure #_ for labelling) | |||
2. Step 2: Use pipettes to dispense 50 μL of the PCR reaction mixes into each of the 8 tubes. | |||
3. Step 3: Using different disposable pipette tips to dispense 50 μL of each DNA sample into their respective tubes | |||
4. Step 4: Be sure to change the pipette tips with each use in order to prevent cross-contamination of the samples | |||
5. Step 5: After all of the tubes contain both the PCR Reaction Mix and the DNA/Primer Mix, place the tubes into the PCR machine and close the lid. | |||
6. Step 6: Make sure the PCR machine is programmed with the correct cycles and temperatures | |||
7. Step 7: Connect the PCR machine to the computer and make sure the power cord is connected to the machine. | |||
8. Step 8: Run the Open PCR software on the computer with the Thermal Cycler program | |||
'''PCR Reaction Mix''' | '''PCR Reaction Mix''' | ||
Each of the 8 tubes will contain 50 μL of the reaction mix. The PCR reaction mixes will contain each of the following: | |||
•Taw DNA Polymerase | |||
•MgCl2 | |||
•dNTP | |||
'''DNA/ primer mix''' | '''DNA/ primer mix''' | ||
In addition to the PCR reaction mix, each tube will contain 50 μL of the following: | |||
•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. | |||
Revision as of 10:37, 23 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 (part 6), 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 will not work when the white wire is disconnected. Test Run (Write the date you first tested Open PCR and your experience(s) with the machine)
ProtocolsThermal Cycler Program
1. Step 1: Label each tube with its respective contents to prevent misdiagnosing patients. (Refer to Figure #_ for labelling) 2. Step 2: Use pipettes to dispense 50 μL of the PCR reaction mixes into each of the 8 tubes. 3. Step 3: Using different disposable pipette tips to dispense 50 μL of each DNA sample into their respective tubes 4. Step 4: Be sure to change the pipette tips with each use in order to prevent cross-contamination of the samples 5. Step 5: After all of the tubes contain both the PCR Reaction Mix and the DNA/Primer Mix, place the tubes into the PCR machine and close the lid. 6. Step 6: Make sure the PCR machine is programmed with the correct cycles and temperatures 7. Step 7: Connect the PCR machine to the computer and make sure the power cord is connected to the machine. 8. Step 8: Run the Open PCR software on the computer with the Thermal Cycler program
PCR Reaction Mix Each of the 8 tubes will contain 50 μL of the reaction mix. The PCR reaction mixes will contain each of the following: •Taw DNA Polymerase •MgCl2 •dNTP
DNA/ primer mix In addition to the PCR reaction mix, each tube will contain 50 μL of the following: •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?
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