Name: Cindy Wong Biomedical Engineering Freshman, Researcher
Name: student Role(s)
LAB 5 WRITE-UP
PCR Reaction Report
Report
The pipetting for this lab was very easy overall, after a few cycles of the pipetting, it became very easy and it was almost like muscle memory.The pre-lab reading aided greatly in understanding the use of lab equipment specifically in the use of the micropipets. The difference between the first stop and the second stop on the micropipetting is the first stop is used to suck up the liquid and the second stop of the pipette is used to shoot the liquid out of the tip of the pipette.The final reactions of the PCR had the same amount of liquid, however the concentration of the DNA within the liquid increased, the actual volume of the liquid did not increase.The tubes appeared to be clean and dry with no visible signs of contamination. We did not have to change our labeling scheme since there were no complications that would have required such an action.
Fluorimeter Procedure
Imaging set-up
To set up our device, a Samsung Galaxy s7, to capture images from the fluorimeter, we had to first set up the camera. The camera's settings were changed to have a film speed, or ISO of 800. Also, the camera flash was inactivated, white balance was changed to auto, exposure and saturation were at its highest setting, and contrast was at its lowest setting. placed the phone on a given smart phone cradle. The smart phone cradle should be at a right angle and positioned where the camera would take images sideways. Next, set the cradle exactly four centimeters away from the drop. From there, pictures can be taken from the fluorimeter.
Placing Samples onto the Fluorimeter
Set the micropipette to 160 microliters and drop 160 microliters of water on the slide. Make sure to position it in the middle first two rows to ensure that the drop looks like a beach ball.
Use a smartphone that has been calibrated to its necessary settings, take three consecutive pictures of the drop.
Dispose of the tip and tray
Set the micropipette to 80 microliters and drop 80 microliters of SYBR Green I in the middle first two rows to ensure that the drop looks like a beach ball.
Add 80 microliters of calf thymus (or water blank)
Adjust the blue LED light to focus on the drop in the middle
Use a smartphone that has been calibrated to its necessary settings, take three consecutive pictures of the drop
Remove the 160 microliter of sample from the surface using a micropipettor
Repeat steps 4-8 for the other concentrations of Calf Thymas DNA
Patient sample demonstrates a concentration reading that is closer to the negative control than the positive control while remaining consistent.
Patient 36290: Inconclusive
Patient sample demonstrates a concentration reading that is inconsistent with data varying greatly depending on different trials yielding inconclusive results.