BME100 f2018:Group6 T1030 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 ReportDuring the first few beginning attempts, a couple of group members were focusing on pipetting which kept the lab going at a fairly steady pace, however, as time went on, another member had been included in order to create a system that was faster, helping the entire process flow smoother. It began to get a little troublesome however to constantly switch from 80 mL to 160 mL in order to clean up the DNA sample/SYBR Green 1 mix, but added to the experience of being able to use the pipette better. Being provided with the pre-lab reading had greatly assisted in preparing the group for how the lab will be carried out and what to expect. The pre-lab reading had as well offered some more insight into what we were working with/ handling in regards to SYBR Green 1 and why it's a more popular dye to work with when -as stated in the reading- looked at from a "green chemistry" perspective. Proper utilization of the pipette due to experience helped smooth out the experience and allowed more time to comprehend what the interaction of the SYBER Green 1 and DNA samples meant when either the SYBER Green 1 wasn't as fluorescent or had not shown up at all. The group understood that the first stop in the pipettor is used in order to discharge the amount of liquid that it was being held to while the second stop is to be used as an extra force to make sure that all the liquid has been removed. During the pipetting process there were a few times when the second stop was used when making sure either all the SYBER Green 1 was used or the DNA samples. From what could be observed there was very little, if not nothing left in the tubes. During the entire process group members pipetting made sure to keep the pipette at 80mL in order to be sure that the correct amount of each sample and SYBER Green 1 was used; being able to use the pipette's second stop had also ensured the amount of liquid used was all 80 mL. The labeling scheme in regards to taking 3 images of the drops, had "changed," that in order to separate them, an arbitrarily taken image had been placed in-between. Fluorimeter ProcedureImaging set-up The blue LED slide holder was first set down, a slide with a water droplet was placed inside. Using the phone stand and an iPhone, the stand and the slide holder was arranged with the slide holder on top of some plastic boxes so that the water droplet was clearly in view. To get a focused, but close up, image, the base of the stand was set up 10cm away from the slide, and the zoom function on the iPhone's camera was used to get a larger image of the drop so the color could be better analyzed. Then, the black box provided was carefully placed over the setup, and then slid forward so that the back wall of the box just touched the slide holder without pushing it forward--this put the sample as far back in the box as possible, which helped to block out most of the light. The iPhone was set with the recommended settings, with maximum exposure, ISO, and saturation, and minimum contrast. The iPhone used a light for the timer countdown, so the photo was manually taken instead. Each photo was taken with the lid of the black box closed as much as possible. Between trials, the entire box was picked up and placed to the side, while the phone cradle and slide holder were carefully left untouched. For each new trial, once the old drop was cleaned up and the new one properly placed, the black box was placed down exactly as it had been--with the back edge carefully put so it was just touching the slide holder. Placing Samples onto the Fluorimeter
Data Collection and AnalysisImages of High, Low, and Zero Calf Thymus DNA Calibrator Mean Values
Calibration curves Images of Our PCR Negative and Positive Controls Positive Control Negative Control
Note: the G6 1 tubes are Patient 1's samples, with the patient ID 23643. The G6 2 tubes are Patient 2's samples, with the patient ID 47311.
PCR Results: Summary
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