BME103:T130 Group 9 l2: Difference between revisions
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Now collection of the data may begin. This is done first by making sure that the experiment worked using the calf thymus DNA as a standard and then measuring the amount of green light from the added dye there is in order to determine the concentration of DNA:<br> | Now collection of the data may begin. This is done first by making sure that the experiment worked using the calf thymus DNA as a standard and then measuring the amount of green light from the added dye there is in order to determine the concentration of DNA:<br> | ||
1. Place a water droplet on the slide in the middle hole.<br> | 1. Place a water droplet on the slide in the middle hole.<br> | ||
2. | 2. Gently add the SYBR Green dye droplet to the water droplet. Place this in the box with the light on and take a picture. This will be the baseline to determine the concentration of pixels in ImageJ when there is no DNA present. <br> | ||
3. Email this photo to a computer with ImageJ and upload it into the program. Split this image into the three component colors: blue, green, and red. Using only the image that shows green, place a circle around the droplet to determine the amount of green in the drop. Make a similar circle on an area of the image that is pitch black. Subtract the background density from the image density to determine the actual density. <br> | |||
4. Repeat steps 2 and 3 but add a droplet of calf thymus to the water droplet as well. This will ensure that the fluorimeter is working properly. Record the actual density of the pixels. This will become important for calculating the concentration of DNA. In order to calculate this, take the actual density and divide it by the calf thymus density and multiply by 2 (since the calf thymus had 2 μL of DNA).<br> | |||
5. Repeat steps 2 through 4 for each sample, including positive and negative controls, and for the three samples from each patient. If the concentration of DNA is similar to that of the calf thymus, this indicates that the person likely has the mutation associated with colon, rectal and/or pancreatic cancer.<br><br> | |||
'''Use of OpenPCR and Fluorimeter for More Cancer Markers''' | |||
==Research and Development== | ==Research and Development== |
Revision as of 20:10, 19 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
The following materials will need to be supplied by the user:
PCR Protocol In order to perform PCR, the samples must first be prepared. This is done by adhering to the following steps: Now that the samples have been prepared, the PCR machine must then be configured to the specific cycling and temperature needs of your experiment. In this case, the instructions for carrying out PCR in one manner are as follows:
Now collection of the data may begin. This is done first by making sure that the experiment worked using the calf thymus DNA as a standard and then measuring the amount of green light from the added dye there is in order to determine the concentration of DNA: Research and DevelopmentBackground on Disease Markers
Primer Design
Illustration
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