BME103:T930 Group 10 l2: Difference between revisions
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'''PCR Protocol''' | '''PCR Protocol''' | ||
1. Label Eppendorf Tubes with 1-17, 2 for positive and negative controls and the rest for the patients DNA. Each tube should contain 50 microliters of one of the following substances: <br> | |||
Positive Control, Negative Control, Patients 1-5 each with Replicates 1-3 <br> | |||
2. Use properly labeled micropipette to place 17 samples into 17 properly labeled Eppendorf tubes already containing 50 microliters GoTaq mix. To see contents of GoTaq mix, see materials.<br> | |||
3. Place 17 Eppendorf tubes in Open PCR. This PCR machine is made of steel in order to decrease safety hazards, and is able to hold more Eppendorf tubes. Therefore, all 17 Eppendorf tubes fit in this PCR. The top of the PCR machine is snaps shut and can be clipped down to be less dangerous. <br> | |||
4. To make up for the more Eppendorf tubes in the PCR machine, each cycle is increased by 2 degrees Celsius, making the Thermal Cycler program: <br> | |||
Stage One: 1 cycle, 95 degrees Celsius for 180 seconds<br> | |||
Stage Two: 35 cycles, 95 degrees Celsius for 30 seconds, 57 degrees for 30 seconds, 72 degrees Celsius for 30 seconds<br> | |||
Stage Three: 72 degrees Celsius for 180 seconds<br> | |||
Final Hold: 4 degrees Celsius<br> | |||
5. When PCR is complete, proceed to DNA Measurement Protocol<br> | |||
'''DNA Measurement Protocol''' | '''DNA Measurement Protocol''' | ||
==Research and Development== | ==Research and Development== |
Revision as of 10:55, 15 November 2012
BME 103 Fall 2012 | Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help | |||||
OUR TEAMLAB 2 WRITE-UPThermal Cycler EngineeringOur re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski.
Key Features
Instructions
ProtocolsMaterials
PCR Protocol 1. Label Eppendorf Tubes with 1-17, 2 for positive and negative controls and the rest for the patients DNA. Each tube should contain 50 microliters of one of the following substances: DNA Measurement Protocol Research and DevelopmentBackground on Disease Markers
Primer Design
Illustration
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