BME100 f2015:Group10 8amL5

BME 100 Fall 2015	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 Error creating thumbnail: File missing Name: Jose Luis Rivera Role(s): Micropipetting, Fluorimeter Procedure, and Data Analysis Error creating thumbnail: File missing Name: Amity Jackson Role(s): Data Collection and Analysis Error creating thumbnail: File missing Name: Jarrett Eshima Role(s): Assisted with Micropipetting, Fluorimeter Procedure, and Data Analysis Error creating thumbnail: File missing Name: Sheldon Cummings Role(s): Data Collection and Analysis Error creating thumbnail: File missing Name: Diba Pourazar Role(s): Collecting materials and Data Collection and Analysis Error creating thumbnail: File missing Name: Katarina Junio Role(s): Micropipetting, Fluorimeter Procedure, and Data Analysis, PCR Reaction Report LAB 5 WRITE-UP PCR Reaction Report The pre-lab readings and activities were helpful in teaching us how to micropipette. It reiterated the steps in order to instill in our heads exactly how to micropipette, since PCR deals with very precise samples. We learned the difference between the first and second stop on the pipettor, in that the first stop is used for collecting the sample, while the second stop is used for releasing the sample. In terms of actually collecting the samples, the final reactions had the same amount of liquid, with the exception of our positive control (G10+). That reaction had slightly less liquid, due to the positive control DNA sample having less than 50 μL in the tube. There was also a little bit of liquid left over in some of the PCR reaction mix, but it was only very little, being just a single drop at the very bottom of the tube. We also did not have to change our labeling scheme. Fluorimeter Procedure Smart Phone Camera Settings Type of Smartphone: Samsung Galaxy S5 Flash: Off ISO setting: Auto White Balance: Auto Exposure: Max Value +2.0 Saturation: N/A Contrast: N/A Camera set-up Place the phone in the phone cradle. Adjust camera setting as listed above. Adjust height so that the droplet is leveled with the camera. Measure the distance from the camera to the droplet and keep it constant. Distance between the smart phone cradle and drop = 9 cm Placing Samples onto the Fluorimeter Place the slide in the Fluorimeter smooth side down. Pipette an 80 uL drop of SYBR Green 1 solution onto the slide. Pipette an 80 uL drop of sample solutions on top of the SYBR Green 1 drop. The drop should rest in the middle of two circles. The solution should not be placed on a previously used portion of the slide. Adjust the slide so that the blue light lies on the drop of solution. Adjust the smartphone so that it is 9 cm from the drop to the camera. Set a timer on the phone camera and close the flap to allow for a dark environment for the picture. Pipette the liquid and discard it in the designated area. Move the slide into the next position and repeat. Data Collection and Analysis Images of High, Low, and Zero Calf Thymus DNA 5 μg/mL sample Error creating thumbnail: File missing 0.5 μg/mL sample Error creating thumbnail: File missing Zero DNA Error creating thumbnail: File missing Calibrator Mean Values Initial Concentration of 2X Calf Thymus DNA solution (micrograms/mL) Final DNA concentration in SYBR Green I solution (µg/mL) Sample Number RAWINTDEN DROP - BACKGROUND MEAN Standard Deviation Image 1 Image 2 Image 3 5 2.5 C-1 1680573 1771664 1706630 1719622.333 46914.74304 2 1 C-2 1206635 2028140 1979381 1738052 460865.9034 1 0.5 C-3 1363342 1368474 1397853 1376556.333 18621.09998 0.5 0.25 C-4 855344 982490 920911 919581.6667 63583.42295 0.25 0.125 C-5 687183 698420 682514 689372.3333 8175.884927 0 0 C-6 410820 382336 460761 417972.3333 39698.70351 Calibration curves Error creating thumbnail: File missing Error creating thumbnail: File missing Images of Our PCR Negative and Positive Controls Negative Control Positive Control PCR Results: PCR concentrations solved PCR Product TUBE LABEL MEAN (of RAWINTDEN DROP - BACKGROUND) PCR Product Concentration (µg/mL) (Step 5 calculation) Total Dilution Initial PCR Product Concentration (µg/mL) (Step 6 calculation) G10 + 1561781 1.568894033 12 18.82672839 G10 - 554868 0.02326170415 12 0.2791404498 G10 1-1 2543619.33 3.076036223 12 36.91243468 G10 1-2 1945548.66 2.157985347 12 25.89582416 G10 1-3 1881616.33 2.059847895 12 24.71817474 G10 2-1 744948.66 0.3150394577 12 3.780473492 G10 2-2 1017554.66 0.733495319 12 8.801943827 G10 2-3 708351.66 0.258862304 12 3.106347648 PCR Results: Summary Our positive control PCR result was 18.82672839 μg/mL Our negative control PCR result was 0.2791404498 μg/mL Observed results Patient 54926: Since this patient was positive, the images portrayed a droplet that glowed green. The average quantitative description (μg/mL) of this patient was 29.18 μg/mL. Patient 57766: Since this patient was negative, the images portrayed a droplet that was translucent. The average quantitative description (μg/mL) of this patient was 5.23 μg/mL. Conclusions Patient 54926 : The average for this patient was 29.18 ug/mL which is much closer to the positive control, 18.83 ug/mL, than the negative control, 0.279 ug/mL. Therefore, the patient can be concluded as positive because of the quantitative analysis. Patient 57766 : The average for this patient was 5.23 ug/mL which is much closer to the negative control, 0.279 ug/mL, than the positive control, 18.83 ug/mL. Therefore, the patient can be concluded as negative because of the quantitative analysis.