BME103:T930 Group 10 l2: Difference between revisions
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'''PCR Protocol''' | '''PCR Protocol'''<br> | ||
BE CAREFUL NOT TO CROSS CONTAMINATE!!<br> | BE CAREFUL NOT TO CROSS CONTAMINATE!!<br> | ||
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1. With permanent marker, clearly number micropipettes. With permanent marker, number Eppendorf tubes at the top. There should be 17 labeled micropipettes and 17 Eppendorf tubes. <br> | 1. With permanent marker, clearly number micropipettes. With permanent marker, number Eppendorf tubes at the top. There should be 17 labeled micropipettes and 17 Eppendorf tubes. <br> | ||
2. Transfer each sample from PCR Eppendorf tubes to Eppendorf tubes containing 400 microliters of buffer. Get all of the sample into the Eppendorf | 2. Transfer each sample from PCR Eppendorf tubes to labeled (1-17) Eppendorf tubes containing 400 microliters of buffer. Get all of the sample into the Eppendorf tubes. <br> | ||
3. Take specially labeled Eppendorf tube (18) with SYBR GREEN I and using specially labeled micropipette (18) place 2 drops on first two centered drops on fluorimeter slide. These dots are further apart on this fluorimeter slide in order to avoid contamination.<br> | 3. Take specially labeled Eppendorf tube (18) with SYBR GREEN I and using specially labeled micropipette (18) place 2 drops on first two centered drops on fluorimeter slide. These dots are further apart on this fluorimeter slide in order to avoid contamination.<br> | ||
4. Take diluted sample from Eppendorf | 4. Take diluted sample from labeled Eppendorf tubes and place 2 drops on top of sample of SYBR GREENI drop.<br> | ||
5. Turn light on, place beam so that it is passing through the drop.<br> | 5. Turn light on, place beam so that it is passing through the drop.<br> | ||
6. Take picture with built in camera to avoid variability on smart phone camera settings. <br> | 6. Take picture with built in camera to avoid variability on smart phone camera settings. <br> | ||
7. Use micropipette properly with black strip for waste to discard sample. <br> | 7. Use micropipette properly labeled with black strip for waste to discard sample. <br> | ||
8. Repeat 3-7 for all samples (Eppendorf tubes 1-17), the DNA of rabbit eyeball (Eppendorf tube 19), and scintillation vial. <br> | 8. Repeat 3-7 for all samples (Eppendorf tubes 1-17), the DNA of rabbit eyeball (Eppendorf tube 19), and scintillation vial. <br> | ||
9. Generate pictures in ImageJ to analyze results. <br> | 9. Generate pictures in ImageJ to analyze results. <br> |
Revision as of 11:23, 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 For PCR Protocol
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