BME100 f2016:Group8 W8AM L5
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LAB 5 WRITE-UP
PCR Reaction Report
Pipetting the samples into the PCR reaction was not a difficult task to accomplish. The first step in pipetting is to set the volume to 250 uL. This is done by twisting the top of the pipette until it reaches this measurement. Then we attach a micro pipette tip to the pipette by pressing down on the pipette tip. Then one has to press down on the plunger on the top of the pipette until it reaches the first stop and no further, and hold the plunger down as the pipette is lowered into the sample. Then the plunger is slowly released to its start position, and as this is happening, the pipette tip sucks up the liquid. After waiting one or two seconds and making sure the tip has no air bubbles, the tip is slowly and vertically pulled out of the tube and lowered into the PCR reaction tube that will hold our samples when we put it in the PCR machine. Then the plunger is pressed down to the second stop so that all of the sample is dispelled into the PCR reaction tube, and after waiting one or two seconds to make sure all the liquid is dispelled into the tube, the tip is removed from the tube and the plunger is slowly released. After the sample is in the PCR reaction tube and the tube is closed, the pipette tip is ejected from the pipette by pressing down on the eject plunger on the side of the pipette into a disposal bag. Overall, our experience with pipetting went well as we followed the instructions and knew what we were doing. The pre-lab reading did help because it helped give us a basic understanding of how the pipette worked and how pipette technique works. The pipette videos also helped us understand the difference between the first and second stop on the pipettor, but we also had previous knowledge on how the pipettor worked and proper pipetting technique. The final reactions did have the same amount of liquid because the precise amount of 80 uL of the sample and the PCR reaction mixtures were added into the reaction tube with the pipettor. However, there could've been errors made, as some sample pipetting could have had some air bubbles in the tips causing some inaccurate amount of liquid. There was no concentrated sample solution left in the sample tubes, but there was some diluted sample patient solution left in the tubes that were pipetted and some liquid left in the tubes that held the PCR reaction mix. We didn't change any of the labeling scheme, as all the tubes were properly labeled with the patient samples, positive control, and negative control.
1) Acquire a smartphone
2) Open camera app, set the timer to three seconds, and turn the camera flash off if possible.
3) Place smartphone in cradle, and adjust height if needed.
4) Camera set up: vertically align camera in cradle so that camera lens is at the same level as where the drop will be placed.
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
PCR Results: Summary