BME103 s2013:T900 Group7 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: Samantha Barker Name: Carissa Henriksen Name: Amanda Seaney Name: Nathan Scjeuer Name: student LAB 2 WRITE-UP Background Information SYBR Green Dye SAMANTHA SYBR Green Dye is a membrane permeable nucleic acid dye used in molecular biology labs. The green dye can be placed in a sample with possible DNA and will glow a bright green if DNA is present. This is the most common use of the dye and helps qualitatively analyze if DNA is present in a substance. Although the DNA used in our samples was double stranded DNA, the SYBR Green Dye can also be used to detect single stranded DNA because it has such a high affinity. Although it was placed in the samples after being in the PCR machine, it also can be placed inside a PCR mixture and this allows the dye to bind to the single stranded DNA before it is synthesized by the Taq Polymerase. However, the result is the same. Either way it binds to DNA and glows when DNA is present. Single-Drop Fluorimeter [A description of the single-drop fluorimeter device. Add a PHOTO for bonus points] How the Fluorescence Technique Works SAMANTHA [In your own words, a summary of the information from page 9 of the worksheet] Procedure Smart Phone Camera Settings iPhone 4s Flash: Off Mode ISO setting: N/A White Balance: N/A Exposure: N/A Saturation: N/A Contrast: N/A Calibration The distance between the smart phone cradle and the drop is approximately 10 cm. Solutions Used for Calibration Calf Thymus DNA solution concentration (micro/mL) Volume of the 2X DNA solution (μL) Volume of the SYBR GREEN I Dye solution (μL) Final 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 One must align the slide with the blue LED. One should place 80 μL of SYBR GREEN in between two wells. It is necessary for one to now add 80 μL of DNA sample in the drop that is formed from step #2. One must cover this created sample from light in order to visualize the florescence. Data Analysis Representative Images of Samples Image J Values for All Samples DNA Concentration Area Density 0 13856 225126 0 13856 51984 0 13856 68928 0.25 13856 199688 .025 13856 190251 .025 13856 273555 0.5 13856 405395 0.5 13856 482091 0.5 13856 636335 1 13856 1335751 1 13856 1274172 1 13856 1105412 2 13856 1895959 2 13856 1372946 2 13856 1769988 5 13856 1728032 5 13856 2042370 5 13856 1918408 Fitting a Straight Line