BME103 s2013:T900 Group6 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: Dale Franco L. Caagbay - Protocol Name: Cyril Wassef - Background Information Name: Emmanuel Casildo - Background Information Name: student Name: student Name: student LAB 2 WRITE-UP Background Information SYBR Green Dye SBYR green dye is a nucleic acid-sensitive gel dye that is useful in pinpointing double stranded DNA in samples within open PCR machines and other utilities such as the single-drop fluorimeter. Single-Drop Fluorimeter The single-drop fluorimeter is a fluorescence-based DNA detection device that aids in bringing to light PCR-Amplified DNA. The DNA sample is placed on a slide upon the machine and shined with a blue LED light. When the light hits the sample, the sample's DNA emits a dim green glow. From there, under a controlled, light-limited environment, a camera (specifically a camera on a phone) can be used to capture the green light. 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 Iphone 5 Flash: None ISO setting: NA White Balance: NA Exposure: NA Saturation: NA Contrast: NA Calibration Photo of Fluorimeter Distance between the smart phone cradle and drop = 8cm Solutions Used for Calibration Calf Thymus DNA solution concentration (microg/mL) Volume of the 2X DNA solution (uL) Volume of the SYBR GREEN I Dye Solution (uL) Final DNA concentration in PicoGreen Assay (ng/mL) 0 80 80 blank .25 80 80 .125 .5 80 80 .25 1 80 80 .5 2 80 80 1 5 80 80 2.5 Placing Samples onto the Fluorimeter Obtain waterproof slides, a Micropipette, and vials of various DNA samples along with a 200 ml vial of SYBR Green solution. Insert slide into fluorimeter Turn on blue fluorescent light and make sure that the beam is shined between the first two rows of the dotted slide. Take 80 micro-liters of SYBR Green solution into micropipette Release all of the 80 micro-liters of SYBR Green fluid onto the slide, then ensure that the beam of light hits the droplet. Take another 80 micro-liters of DNA Sample and extract into the SYBR Green droplet. Make sure the drops form a ball and doesn't spread around making individual droplets all over the slide. After the photo shoot is complete, calibrate the micropipette to 160 micro-liters and extract all of the sample on the slide. Repeat the same steps for the additional DNA Samples. Clean Up Workstation 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]