BME103 s2013:T900 Group2 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: Andy Son Mitch Riggs Open PCR Machine Engineer/ Team Leader Name: William Scott Name: Joe Sansone Name: student LAB 2 WRITE-UP Background Information SYBR Green Dye SYBR Green dye is a dye used as an indicator in molecular biology. It emits green light when bound to double helix DNA and illuminated with blue light. Source: http://en.wikipedia.org/wiki/SYBR_Green_I 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: No flash was used ISO setting: White Balance: Exposure: Saturation: Contrast: Calibration [Describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points.] Distance between the smart phone cradle and drop = 7cm Solutions Used for Calibration Calf Thymus DNA Solution (microg/mL) Volume 2X DNA Solution (uL) Volume SYBR GREEN I Solution (uL) Fina DNA concentration in PicoGreen Assay (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 0.125 0 80 80 blank Placing Samples onto the Fluorimeter Step 1: A micro pipette was obtained and 80uL of the SYBR GREEN I was dispensed in the center of the first two rows of the slide. Once done 80uL of the desired solution (calf thymus at various concentrations or just water) was added to the mixture on the slide. Step two: After the desired mixture was placed onto the slide it was then positioned so that the droplet was centered with the small opening on the apparatus where the blue LED light originates from. Step three: After collecting the qualitative data with picture captures, the sample was removed from the slide and discarded appropriately. Then the above steps were repeated for all concentrations of DNA making sure not cross contaminate the mixtures by using different slide rows for each trial. Step four: Once all experimental trials were done the work station was cleaned by discarding the slides in the appropriate sharps container as well as dispensing the DNA solutions appropriately. The data was compiled and tabulated for further analysis. 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] 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 [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]