BME100 s2014:T Group8 L5: Difference between revisions
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'''PCR Results Summary'''<br> | '''PCR Results Summary'''<br> | ||
A Polymerase Chain Reaction (PCR) was completed for 8 DNA samples--3 DNA samples from each of two patients and a positive and a negative control sample. After the reactions and | A Polymerase Chain Reaction (PCR) was completed for 8 DNA samples--3 DNA samples from each of two patients and a positive and a negative control sample. After the reactions proceeded and cooled down, the samples were tested on a fluorimeter for amplification of DNA using SYBR Green I as a stain. The controls were necessary to ensure amplification results for the patients' samples had a comparative standard and the results were due to the procedure and not an external affect. Using a known concentration of DNA from the calibration phase, a standard curve was created that was a basis for conversion from imageJ INTEN values to DNA concentrations for the PCR samples. <br> | ||
Your positive control PCR result was 90.744 μg/mL <br> | Your positive control PCR result was 90.744 μg/mL <br> |
Revision as of 01:25, 17 April 2014
BME 100 Spring 2014 | 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 TEAM
LAB 5 WRITE-UPBackground InformationSYBR Green Dye
The way the fluorescence works is by utilizing a green dye that activates only under the presence of double stranded DNA (or dsDNA).
ProcedureSmart Phone Camera Settings
The camera phone was set up in the cradle with an angle to view the side of the drop. The fluorimeter was raised about 1 1/4 inches to accomodate for the phone height.
The phone was removed from this image to simplify and completely show distance setup.
Solutions Used for Calibration
1. Turn on the flourimeter. Place the super hydrophobic slide on the flourimeter. On the hydrophobic side of the flourimeter, place a 80 microliter drop of SYBR GREEN I on the middle rows of the slide. 2. Now on the sphere of dye that is on the slide, add 80 microliter of one of the solutions being tested. This will now react or not. 3. Next the flourimeter will shine a blue light, align the slide so that the drop sample is in the middle of this light. 4. Acquire a box that can cover the flourimeter in order to create a dark environment. Place a smartphone in the cradle, and use a timer on the camera to take a picture of the drop. Once the timer starts quickly put the box over the whole setup, so that outside light does not interfere.
Data AnalysisRepresentative Images of Samples High DNA - using green channel
Fitting a Straight Line Intends values for each Calf Thymus DNA cocnentration. PCR Results Summary A Polymerase Chain Reaction (PCR) was completed for 8 DNA samples--3 DNA samples from each of two patients and a positive and a negative control sample. After the reactions proceeded and cooled down, the samples were tested on a fluorimeter for amplification of DNA using SYBR Green I as a stain. The controls were necessary to ensure amplification results for the patients' samples had a comparative standard and the results were due to the procedure and not an external affect. Using a known concentration of DNA from the calibration phase, a standard curve was created that was a basis for conversion from imageJ INTEN values to DNA concentrations for the PCR samples. Your positive control PCR result was 90.744 μg/mL Write-in each patient ID and give both a qualitative (what the images looked like) and a quantitative description (μg/mL) of what you observed
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