BME100 f2018:Group7 T0800 L5
BME 100 Fall 2018 | 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-UPPCR Reaction ReportWe gathered all of the materials in stages, first we were given eight empty PCR tubes and an empty PCR tube rack. Then we were given PCR mix, a positive and negative control, and finally our patient samples. We were advised to handle the materials carefully with disposable gloves and lab coats on. We started pipetting be calibrating the micropipette to 50 microliters. We began placing the PCR mix into each PCR tube, switching people after every two tubes so that each group member could gain experience with performing the experiment. After a little practice and watching the pre-lab video that demonstrated how to pipette, the application was relatively easy. Through brief exposure to using the pipette we were able to distinguish the purposes of the first and second stops on the pipette, which made transferring the liquid easier. We then added the controls and each patient mixture to each PCR tube accordingly. We did not have the same exact amount of liquid present in the final reactions. This is likely due to consistently switching who operated the pipette, because we likely applied different pressures when pressing on the first and second stops. The volume discrepancies could also have occurred due to the placement of the pipette as it gathered the liquid. For instance, we would get slightly different volumes if we placed the pipette tip all the way at the bottom of the tubes, versus somewhere in the middle of the liquid solution. Consequently, there was some liquid left in the tubes due to the different volumes of liquid gathered. We didn't have to change our labeling scheme, but we made sure to follow the procedure by starting with the positive and negative controls, and then going through the 6 replicates of the two patients as found in Part A. Fluorimeter ProcedureImaging set-up The device that was used for this part of the experiment was an iPhone. The first thing we did was to make sure our iPhone was placed in the appropriate position, making sure that the camera lens was consistently lined up with the slide. This ensured having a significant change in the distance between the iPhone cradle and the droplet on the slide. The iPhone was placed on the cradle and a ruler was used to measure the distance. We placed the iPhone cradle 9cm away from the slide, and then checked that the camera was focused on the drop, to prevent blurry pictures. Despite this effort, we had blurry images for the calibration component of data, and so to perform the rest of the calculations as proof of principle, we had to zoom in on the images to get a clearer representation of them for processing in imageJ. Placing Samples onto the Fluorimeter
Data Collection and AnalysisImages of High, Low, and Zero Calf Thymus DNA
Images of Our PCR Negative and Positive Controls PCR Results: PCR concentrations solved
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