BME100 f2016:Group10 W1030AM L5

BME 100 Fall 2016	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 Amy Polanecki Tariq Madni Kyle Hull Andrea Hnatievych Neaco Fox LAB 5 WRITE-UP PCR Reaction Report The pre-lab reading was very interactive and informative. For those who have never used a micropipette, the pre-lab information was beneficial. The first stop on the pipette is used to collect the sample that is being transferred and the second stop is used to release the sample liquid. The final reaction will have the same amount of liquid. There was no liquids left in the tubes of the DNA samples and PCR reaction mix. We did not change our labeling scheme, but due to the tubes being very small we could not fit the whole label onto it. We labeled the first tube as G10 + and the second tube as G10-, but for the rest of the tubes we did not add "G10" to the labeling. Rather, we labeled the other 6 tubes as 1-1,1-2,1-3,2-1,2-2, and 2-3. Fluorimeter Procedure Imaging set-up A member in our group provided their iPhone 6s camera in order to take images while the fluorimeter measures the fluorescence of the DNA. This is the transmittance of the wavelength given off by the DNA that is excited. The wavelength of light is exciting the DNA and if it fluoresces then there is double-stranded DNA present. Taking the iPhone 6s, the camera's flash was inactived in order to get a reasonable image of the calibration with the DNA concentration. The iPhone 6s was placed on to a metal card holder and the metal card holder was put on top of a plastic stand at a distance of 4 centimeters. The iPhone 6s was leaning on an angle due to the metal card holder and we wanted to have the camera perpendicular to the fluorimeter. In order to do this, we placed a folded piece of paper behind the iPhone 6s to prop it up. Once our set up was completed, we began capturing images of our DNA. A black box was placed over the fluorimeter and camera in order to ward off excess light. Before closing the box and omitting all light, the iPhone 6s timer was set at 10 seconds in order to allow the camera to focus on the new lighting exposure. The lid of the box was closed and after 10 seconds the camera took the photo. The procedure for capturing the image was repeated three times per DNA sample. Placing Samples onto the Fluorimeter Place a clean slide onto the fluorimeter. Take a micro-pipette and place a disposable tip onto it. Using the micro-pipette, take 80 microliters of a sample of 2X Calf Thymus DNA solution. Transfer the solution onto the fluorimeter and notice that it should create a "beach ball" bubble. Release the solution onto the front half of the slide so the back half of the slide can be used for another test. Remove the disposable tip and toss it into a trash cup. Place another clean, disposable tip onto the micro-pipette. Using the micro-pipette, take 80 microliters of a sample of SYBR GREEN I Dye solution. Release the solution onto the "beach ball" bubble of 2X Calf Thymus DNA solution. Move the slide containing the drop of the two solutions within the fluorimeter. Align the sample with the blue LED light and make sure the light shines through the drop. Set up the camera and place the black box over the fluorimeter and camera. Once the three pictures of the solution are captured, take the micropipette and remove the sample on the fluorimeter. Dispose of the sample in the micro-pipette into the trash cup and remove the disposable tip. Repeat steps 1-12 for the 6 initial concentrations of 2X Calf Thymus DNA solution. Place a clean slide onto the fluorimeter. Take a micro-pipette and place a disposable tip onto it. Using the micro-pipette, take 80 microliters of a sample of 2X DNA solution. Transfer the solution onto the fluorimeter and notice that it should create a "beach ball" bubble. Release the solution onto the front half of the slide so the back half of the slide can be used for another test. Remove the disposable tip and toss it into a trash cup. Place another clean, disposable tip onto the micro-pipette. Using the micro-pipette, take 80 microliters of a sample of SYBR GREEN I Dye solution. Release the solution onto the "beach ball" bubble of 2X DNA solution. Move the slide containing the drop of the two solutions within the fluorimeter. Align the sample with the blue LED light and make sure the light shines through the drop. Set up the camera and place the black box over the fluorimeter and camera. Once the three pictures of the solution are captured, take the micropipette and remove the sample on the fluorimeter. Dispose of the sample in the micro-pipette into the trash cup and remove the disposable tip. Repeat steps 14-25 for the 6 2X DNA solutions. Data Collection and Analysis Images of High, Low, and Zero Calf Thymus DNA Calibrator Mean Values TABLE GOES HERE Calibration curves Images of Our PCR Negative and Positive Controls PCR Results: PCR concentrations solved TABLE GOES HERE PCR Results: Summary Our positive control PCR result was ____ μg/mL Our negative control PCR result was ____ μg/mL Observed results Patient 77434 : Patient 77218 : Conclusions Patient 77434 : Patient 77218 :