BME100 f2015:Group12 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: Krystal Schmitt Error creating thumbnail: File missing Name: Ladan Kamali Sarvestani Error creating thumbnail: File missing Name: Nathan McAdams Error creating thumbnail: File missing Name: Ashwin Krishnadoss Error creating thumbnail: File missing Name: Fangchi Shao Error creating thumbnail: File missing Name: Matthew Levins LAB 5 WRITE-UP PCR Reaction Report In today's Lab, we used micropipets to transfers samples into test tubes. Our prelab this week went over how to properly use the micropipets, specifically with differentiating between the first stop and the second stop on the pipet and how to utilize them (first stop for pulling up sample and second stop for discarding sample.) The prelab was extremely helpful for the students who had no prior experience with the micropipets and it was also a good refresher for those who hadn't used them in a while. Conducting the lab was very successful; however, there was one instant where some fluid got sucked back up into the pipet after discharge and we were unable to get the rest out (a very little amount.) Fluorimeter Procedure Smart Phone Camera Settings Type of Smartphone: Samsung edge Flash: Off ISO setting: N/A White Balance: N/A Exposure: N/A Saturation: N/A Contrast: N/A Camera set-up Distance between the smart phone cradle and drop = 15cm Placing Samples onto the Fluorimeter First, place a 160 microliter drop of H20 on a slide and place it in the fluorimeter with the black box covering the apparatus and the blue beam shining through the drop Using a smart phone, capture a profile picture of the drop (making sure the set up was good enough to continue) Place 80 microliters of SYBR green solution on a slide and add 80 microliters of the 5 micrograms/mL of calf thymus solution Take 3 separate pictures of this drop in the black box with the blue beam shining through it Next, discard the solution from the slide and adjusted it for use of a new drop (or grab an entire new slide if necessary) We repeated steps 3-5 for 2, 1, 0.5, and 0.25 micrograms/mL of calf thymus solution Next, transfer 100 microliters of each patient sample (PCR from last lab) into a buffer solution tube (using a difference tube for each PCR sample) and invert them to mix solutions Using a slide, transfer 80 microliters of SYBR green and 80 microliters of each buffer solution (do each one separately) Take 3 pictures of the drops created on the slides for each SYBR green, PCR mixture Data Collection and Analysis Images of High, Low, and Zero Calf Thymus DNA 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 0.5 0.25 C-1 36672 29037 28290 8382 4638.77063 0.25 0.125 C-2 31743 31879 30522 31381.33 747.3047125 1 0.5 C-3 38563 38902 31748 36404.33 4036.063718 2 1 C-4 40395 39924 35530 38616.33 2683.197781 5 2.5 C-5 48724 42633 42889 44748.67 3445.118334 0 0 C-6 0 0 0 0 0 Calibration curves Images of Our PCR Negative and Positive Controls 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) ─ 2661320 167.3764684 12 2008.51762 y 2675348 168.26241 12 2019.14892 # 2298222 144.4449286 12 1733.339144 ? 2654999 166.9772641 12 2003.727169 + 2715039 170.7691045 12 2049.229254 x 2128944 133.7541367 12 1605.04964 * 2155289 135.4179613 12 1625.015536 ! 2121078 133.2573576 12 1599.088291 PCR Results: Summary Our positive control PCR result was 170.7691045 μg/mL Our negative control PCR result was 167.3764684 μg/mL Observed results Patient 38292 : The image of Patient 38292 appeared to be a drop of water with a green tint. The three trials we conducted produced values of 133.7541367μg/mL, 135.4179613μg/mL, and 133.2573576μg/mL.The mean of the three trials for this patient was 135.1431519μg/mL. Patient 15049 : The image of Patient 15049 appeared to be a drop of water with a slight green tint. The three trials we conducted produced values of 144.4449286μg/mL, 166.9772641μg/mL, and 170.7691045μg/mL. With an average value of 160.7304324μg/mL. Conclusions Patient 38292 : The positive control PCR result was 170.7691045μg/mL. The mean value of the trials for this patient was 135.1431519μg/mL. For this reason, it can be concluded that this patient was negative because of how significantly lower the results were compared to the positive control. Patient 15049 : The positive control PCR result was 170.7691045μg/mL. The mean value of the trials for this patient was 160.7304324μg/mL. For this reason, it can be concluded that this patient was negative because of the low value of the results compared to the positive control. However, this patient was much closer to the positive control compared to the other patient making it more probable that this patient could be positive compared to the other patient.