BME103 s2013:T900 Group5 L2

BME 103 Spring 2013	Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help
OUR TEAM Name: Alexander Oropel Name: Matt McClintock Name: student Name: student Name: student Name: student LAB 2 WRITE-UP Background Information SYBR Green Dye [A short summary describing SYBR green dye] Single-Drop Fluorimeter [A description of the single-drop fluorimeter device. Add a PHOTO for bonus points] How the Fluorescence Technique Works [In your own words, a summary of the information from page 9 of the worksheet] Procedure Smart Phone Camera Settings [The type of smart phone you used and how you adjusted the camera settings, if applicable (see worksheet page 4).] Flash: ISO setting: White Balance: Exposure: Saturation: Contrast: [If you used an additional phone, describe the other type of smart phone you used and how you adjusted the camera settings, if applicable (see worksheet page 4).] Flash: ISO setting: White Balance: Exposure: Saturation: Contrast: Calibration The Camera was set up such that we could get the clearest image with maximum LED light absorption. A light box was placed over the fluorimeter to darken the area. The Smart phone was aligned with the slide to get a sideways picture of the DNA sample. This creates the ideal angle to observe any fluorescent light after the picture is taken. The camera phone is calibrated to view the fluorescent light with little interference from contrast and recording the most light- done by setting the exposure high. The saturation is set to low so that color is not rendered in the photo. The ISO is set to high to maximize focus in the dark setting. With these settings, the fluorescence in the sample is easily observed. Distance between the smart phone cradle and drop = [See worksheet page 5] Solutions Used for Calibration Concentration Calf Thymus (μg/mL) Volume DNA added (μL) Volume SABR GREEN I (μL) Concentration resulting DNA (ng/mL) 5 80 80 2.5 2 80 80 1 1 80 80 0.5 0.5 80 80 0.25 0.25 80 80 .125 0 80 80 blank Placing Samples onto the Fluorimeter 1.) With the Camera set in place we prepared the slide with SYBR Green I, a constant throughout the experiment 2.) 80 μL of 5μg/mL of calf thymus concentration DNA samples were then added to the slide 3.) To take 3 pictures in the the dark with as much of the blue LED light as possible, we used a timer on the camera, set to 3 seconds. 4.) The slide, now containing SYBER Green I and Calf Thymus DNA was then aligned in the cradle such that the light was shining into the DNA. 5.) With the Camera phone, we captured 3 images of the DNA and removed the light box. 6.) That sample was then disposed of with a micropipette 7.) Repeat with 80μL Green SABR I and 80μL of each Calf Thymus Concentration given by the first column in the table above Data Analysis Representative Images of Samples [Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with no DNA] [Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with DNA (positive signal)] Image J Values for All Samples [See worksheet page 5] [Add more rows as needed]Fitting a Straight Line [Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 8] row 1 cell 1 row 1 cell 2 row 1 cell 3 row 1 cell 4 row 1 cell 5 row 2 cell 1 row 2 cell 2 row 2 cell 3 row 2 cell 4 row 2 cell 5 row 3 cell 1 row 3 cell 2 row 3 cell 3 row 3 cell 4 row 2 cell 5