BME100 s2014:W Group14 L5

BME 100 Spring 2014	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: Jose Duran Name: Shawn Garcia Name: Christian Keefer Name: Karthik Puncha Name: Austin Tielke LAB 5 WRITE-UP Background Information SYBR Green Dye [Instructions: A short summary describing SYBR green dye] Single-Drop Fluorimeter [Instructions: A description of the single-drop fluorimeter device. Add a PHOTO for bonus points] How the Fluorescence Technique Works [Instructions: In your own words] Procedure Smart Phone Camera Settings Type of Smartphone: Apple iPhone 5S Flash: Off ISO setting: Auto White Balance: Auto Exposure: High Saturation: High Contrast: Low Calibration The camera was set up by placing the iPhone 5S in the holding compartment of the lab equipment. It was measured to be 6 centimeters away from the origin of the light emission. The settings (stated above) were adjusted; the height of the fluorimeter was adjusted in order to achieve a perfect side-angle perspective of the "beach ball" shaped liquid drop. Distance between the smart phone cradle and drop = 8 centimeters Solutions Used for Calibration Initial Concentration of 2X Calf Thymus DNA solution (μg/mL) Volume of the 2X DNA solution (μL) Volume of the SYBR GREEN I Final DNA concentration in SYBR Green I solution (μg/mL) 5 80 80 2.5 2 80 80 1.0 1 80 80 0.5 0.5 80 80 0.25 0.25 80 80 0.125 0 80 80 0 Placing Samples onto the Fluorimeter Before placing samples onto the fluorimeter, turn on the Blue LED light & have the camera on the smartphone ready. Insert the glass sampling surface, with the smooth side facing down. Position the glass sampling surface so that the light emits between the first two rows of the glass. Place an 80 μL drop of SYBR Green I in the middle of the first two rows. Swap out the pipette tip with a new tip. Place an 80 μL drop of a specifically concentrated Calf Thymus DNA solution onto the top of the previously placed SYBR Green I drop. Every three 160 μL sample drops will vary in concentrated Calf Thymus DNA solution. In order to place a new sample drop, remove the 160 μL solution, move the glass sampling surface to the next two available rows, and repeat the process of placing two new 80 μL drops onto each other (as stated before). Data Analysis Representative Images of Samples Sample w/o DNA Sample w/ DNA Image J Values for All Samples Final DNA Concentration in SYBR Green I Solution (μg/mL) RAWINTDEN of the Drop RAWINTDEN of the Background RAWINTDEN Drop - RAWINTDEN Background 0.5 4421613 187151 4234462 0.5 4521561 175412 4346149 0.5 4159878 179854 3980024 1.0 5904302 195454 5708848 1.0 6048248 187945 5860303 1.0 6248451 198454 6049997 2.5 8558810 212416 8346394 2.5 8821541 205451 8616090 2.5 8258495 215411 8043084 0.0 3201562 167842 3033720 0.0 3158472 154952 3003520 0.0 3584956 151231 3433725 0.125 1124652 165451 959201 0.125 1413586 167851 1245735 0.125 1248412 170245 1078167 0.25 3952152 168672 3783480 0.25 3754546 171541 3583005 0.25 4132142 172164 3959978 Fitting a Straight Line [Instructions: Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 9]