BME100 f2013:W900 Group9 L5

BME 100 Fall 2013	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: Allison Lind Name: Sydney M. Lankford Name: Neel J. Patel Role: Open PCR Machine Testing Name: Callie M. LaMarche Name: Julianna A. Brunner Role: Open PCR Machine Testing LAB 5 WRITE-UP Background Information SYBR Green Dye [Instructions: A short summary describing SYBR green dye] The SYBR green dye has the ability to fluorescene when there is a presence of dsDNA. According to Life Technologies, SYBR dyes are one of the best high sensitivity reagents for staining DNA and RNA. The green dye binds to the DNA, causing the DNA to absorb blue light and reflect green light. Single-Drop Fluorimeter [Instructions: A description of the single-drop fluorimeter device. Add a PHOTO for bonus points] A single-drop fluorimeter device is used to put fluorescene and DNA together on a slide to test for fluoresecene. How the Fluorescence Technique Works [Instructions: In your own words] The fluorescence Technique Works by adding DNA and SYBR Green 1 dye to see how much flourence shines through with the picture. The higher the concentration the more flourence that should be visual when looking at the light shining through the drop. Procedure Smart Phone Camera Settings [Instructions: The type of smart phone you used and how you adjusted the camera settings, if applicable (see worksheet page 4).] Type of Smartphone: iPhone 5 Flash: off ISO setting: auto (could not find option on model used) White Balance: auto (could not find option on model used) Exposure: auto (could not find option on model used) Saturation: auto Contrast: auto Calibration [Instructions: Describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points.] The phone was placed in the cradle at an appropriate angle to view the slide from the side. The fluorimeter was raised in front of the phone to get the right angle. A light box was placed over the set up to provide darkness to take the picture of the light flowing through the DNA and SYBR Green 1. The flourence in the sample was easily seen with higher concentrations of SYBR Green 1. Distance between the smart phone cradle and drop = 7.5 cm [Instructions: See worksheet page 6.] Solutions Used for Calibration [Instructions: See worksheet page 6.] Calf Thymus DNA solution Concentration (microg / mL) Volume of the 2X DNA Solution (μL) Volume of the SYBR GREEN I Dye Solution (μL) Final DNA concentration in SYBR Green I 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 [Instructions: Step one, in your OWN words] [Instructions: Step two, in your own words] [Instructions: Step three, in your own words] [Instructions: Step etc., in your own words] Data Analysis Representative Images of Samples [Instructions: Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with no DNA] [Instructions: 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 [Instructions: See worksheet page 8. To save time on typing a new Wiki table from scratch, use THIS TOOL to auto-generate a Wiki table: Excel-to-Wiki Converter. Copy the headers and values from the Excel spreadsheet you made, paste them into the form field, click submit, copy the Wiki code that the tool generated, and replace TABLE GOES HERE (below) with your auto-generated code.] TABLE GOES HERE Fitting a Straight Line [Instructions: Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 9]