BME103:T930 Group 1 l2: Difference between revisions
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'''DNA Measurement Protocol''' | '''DNA Measurement Protocol''' | ||
'''Image J Procedure''' | |||
1. Search Image J in Google and then download Image J <br> | |||
2. Open Image J then click "file" and click "open" and open the image you want to analyze <br> | |||
3. Once your image is open click "analyze" and then click "set measurements" and check the boxes "area" "integrated density" and "mean gray value" leave the rest of the boxes empty <br> | |||
4. now click "image" then click "color" and then click "split channels" <br> | |||
5. This will split your image into three, you will use the one that is marked as the "green" picture, cancel the others <br> | |||
6. Activate the oval tool <br> | |||
7. draw the best oval you can around the drop and then press the control button+ the "m" key <br> | |||
8. Move the oval over to the background (the black around the picture) and press the control button and the m key again <br> | |||
9. repeat steps for all pictures <br> | |||
10. Save your data in an excel format by clicking "file" and then clicking "save as" then save the file with the name you want <br> | |||
==Research and Development== | ==Research and Development== |
Revision as of 23:06, 28 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
1. You will need a PCR machine and computer. 2. Download the PCR software onto the computer. 3.Thaw the GoTaq Colorless Master Mix at room temperature. Vortex the Master Mix, then spin it briefly in a microcentrifuge to collect the material at the bottom of the tube.
5. If using a cycler without a heated lid, overlay the reaction mix with 1-2 drops of mineral oil to prevent evaporation during thermal cycling. Centrifuge the reaction mix in a microcentrifuge for 5 seconds. 6. Place the reactions in a thermal cycler that has been preheated to 95 degrees Celsius. Perform PCR. What Occurs In the PCR Machine 2. Annealing: perform the reaction about 5 degrees Celsius below the calculated melting temperature of the primers and increasing the temperature in increments of 1°C to the annealing temperature; this should occur anywhere between 30 seconds and 1 minute. 3. Extension: performed between 72-74 degrees Celsius, extension allows 1 minute for every 1 kb of DNA to be amplified; the suggested time for extension is 5 minutes. 4. Refrigeration: refrigerate the tubes at 4 degrees Celsius for several hours; this will minimize the opportunity for DNA polymerase to continue to be active at higher temperatures. 5. Cycle Number: the optimal amplification is 25-30 cycles, but up to 40 may be performed.
Research and DevelopmentBackground on Disease Markers
Primer Design
Illustration
References"GoTaq® Colorless Master Mix (M714) Product Information." GoTaq® Colorless Master Mix Protocol. Promega, 2012. Web. 15 Nov. 2012. <http://www.promega.com/resources/protocols/product-information-sheets/g/gotaq-colorless-master-mix-m714-protocol/>. Hunt, Margaret. "Real Time PCR Tutorial." Real Time PCR Tutorial. University of South Carolina, 10 July 2010. Web. 15 Nov. 2012. <http://pathmicro.med.sc.edu/pcr/realtime-home.htm>. |