OUR TEAM
LAB 5 WRITE-UP
PCR Reaction Report
Fluorimeter Procedure
Imaging set-up Use iphone camera to record pictures of the solution drops
Turn off the flash
Set ISO to 800 or higher
Set white balance to auto
Set exposure to highest setting
Set saturation to the highest setting
Set contrast to the lowest setting
Set the camera timer to 3 seconds
Placing Samples onto the Fluorimeter
- Place 160 microliter drops of H2O betwwen the center rows on a glass test sheet using the pipettor
- Switch on the light in the excitation light
- Turn on and set up camera with correct settings
- Place camera in the cradle and line up the camera to take a profile picture of the solution drop
- Place 80 microliters of SYBR Green I to the solution drop
- Use your timer to take a picture of the solution drop and place the box over the apparatus to prevent light from shining in"
- repeat steps 1-6 for each solution and PCR sample
- record the data
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
High http://i67.tinypic.com/11j0sy1.jpg
Low http://i66.tinypic.com/2vsfd3p.jpg
Zero http://i66.tinypic.com/10cw5r9.jpg
Calibrator Mean Values
http://i68.tinypic.com/14jamnc.jpg
Calibration curves
http://i64.tinypic.com/p2cg4.jpg
http://i63.tinypic.com/24qmsmp.jpg
Images of Our PCR Negative and Positive Controls
Positive Control
http://i68.tinypic.com/2q9xmiw.jpg
Negative Control
http://i64.tinypic.com/23r7cdh.jpg
PCR Results: PCR concentrations solved
http://i67.tinypic.com/a108p4.jpg
http://i68.tinypic.com/fedwyx.jpg
PCR Results: Summary
- Our positive control PCR result was 422.3 μg/mL
- Our negative control PCR result was 185.2 μg/mL
Observed results
- Patient 57027 : The green was very visible on all 3 trials of this patient. We observed 402.1 ug/mL for this patient.
- Patient 97392 :The green dot was not visible in these trials and we observed 223.5 ug/mL for this patient.
Conclusions
- Patient 57027 : This patient's results were very similar to the positive control so we concluded this patient was positive.
- Patient 97392 : This patient's results were very similar to the negative control so we concluded this patient was negative.
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