BME100 s2017:Group7 W1030AM L5

BME 100 Spring 2017	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 Name: Avery Cartwright Name: Angelica Garrido Name: Brianna Lopez Name: Jason Reyes Name: student Name: student LAB 5 WRITE-UP PCR Reaction Report The prelab materials were beneficial in their specificity of demonstration for how exactly micropipetting works and what what the proper technique is. Additionally, having used the previous lab meeting time to prepare for the actual PCR aided in ensuring that the experiment was understood both conceptually and technically and thus able to be executed properly. Because the prelab materials allowed for a firm understanding of micropipetting technique, the difference between the first and second stop on the pipettor were understood and the amount of liquid in each reaction remained constant. Fluorimeter Procedure Imaging set-up The settings for the smartphone camera itself were as follows: flash turned off, ISO set to 800, white balance to auto, exposure and saturation to the highest setting, and contrast to the lowest setting. A black box was placed sideways on the table such that the opening at the top faced outward toward the experimenter in order; this box was used to block out any excess light. The LED light device was placed under the cover the box provided, with the stand for the camera of the smartphone placed 4 cm away from the LED device. The smartphone was then placed inside the stand. Placing Samples onto the Fluorimeter Use the micropipettor to place 160 microliters of the water provided such that it looks like a balloon placed in the first two rows of the hydrophobic side of the slide. Turn on the blue LED setting of the excitation light. Adjust the settings of the smartphone camera so they correspond with the above ramifications. Put the smartphone camera in the provided stand 4 cm away from the LED device and adjust the height so that the camera will be taking pictures at the same height as the drop itself. Take a photo of the water droplet to ensure that the camera is properly placed and set. Use the micropipettor to place 80 microliters of SYBR GREEN I in the middle of the slide such that it once again looks like a balloon; then, add 80 microliters of one of the provided calf thymus solutions. Place the slide in the LED device such that the blue LED light is focused directly on the middle of the drop of the sample. Take three FOCUSED photos of the drop. Remove the box WITHOUT moving the smartphone; if the smartphone does move, ensure that the measurement of the distance between the smartphone and the LED device returns to 4 cm. Remove that 160 microliter drop with the pipettor and then move the slide to the next position. Repeat these steps for all the different concentrations of the calf thymus DNA. Data Collection and Analysis Images of High, Low, and Zero Calf Thymus DNA 5 μg/mL sample 0.5 μg/mL sample 0 μg/mL sample Calibrator Mean Values Calibration curves Images of Our PCR Negative and Positive Controls Positive Control Negative Control PCR Results: PCR concentrations solved PCR Results: Summary Our positive control PCR result was -56.65 μg/mL Our negative control PCR result was 61.64 μg/mL Observed results Patient 91754 : For the 5μg/mL sample, the fluorimeter result had a bright white color. For the 0.5μg/mL, the fluorimeter result had a white hue to it, but was considerably darker than the 5μg/mL. In the 0μg/mL, the fluorimeter result had some white hues to it, but was considerably lass reflective and darker in color than the previous results. Patient 99764 : The results of this patient were similar to those of the previous patient. For the 5μg/mL sample, the fluorimeter result had a bright white color. For the 0.5μg/mL, the fluorimeter result had a white hue to it, but was considerably darker than the 5μg/mL. In the 0μg/mL, the fluorimeter result had some white hues to it, but was considerably lass reflective and darker in color than the previous results. Conclusions Patient 91754 : From the observed result, it can be said that this patient is negative. This is from the observation of having the distinct white coloring from the putting it through the lab fade. This is also is confirmed from this patient results to be more closely related to the negative control than to the positive control. Patient 99764 : In a similar case, this patient was also negative. The white coloring from the process in the beginning slowly drained, resting into a more see through sample. The result from the lab also consistently put it more closely to the negative control result than the positive control result. Gel Electrophoresis Results From the ladder on the left to the right the cells are: positive control, negative control, patient 1 replicate 1, patient 1 replicate 2, patient 1 replicate 3, patient 2 replicate 1, patient 2 replicate 2, patient 2 replicate 3. The patients had a negative result, as shown above in the gel electrophoresis. The only positive that was shown by the gel was the positive control, but in the end the patients had an overall negative result.