BME100 s2014:T Group14 L5: Difference between revisions
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'''How the Fluorescence Technique Works' | '''How the Fluorescence Technique Works''' | ||
First, the flourimeter is set up by turning on the Blue LED and turning on and putting a smart phone on a cradle in front of the flourimeter. From this position, adjust the flourimeter to the height of the cell phone's camera in order to get a picture of the drop sideways and not from any other angle. This can be done by placing books or other materials underneath the flourimeter. After placing the drop of SYBR Green on the slide and adding the DNA to the dye, the dye will bind to the nucletide in the DNA. So, when the SYBR Green is exposed to the blue light, it will start to flouresce, putting a dark container over the reaction will make the flourescence easier to see in the picture taken by your cell phone. | |||
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Revision as of 19:29, 16 April 2014
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
LAB 5 WRITE-UPBackground InformationSYBR Green Dye SYBR Green I is a dye that is used as a nucleic acid stain in molecular biology. The dye binds to DNA and results in a DNA and dye complex that reflects blue light and emits a green light that binds to double stranded DNA. This can be used in some methods of a quantitative polymerase chain reaction. This dye can also work on RNA, because RNA also is made up of nucleic acids. The dye will bind to the RNA and work in the exact way it would when bound to the DNA. SYBR Green I is declared safe to work with and does not have waste disposal issues. However is has the cabable of binding to DNA with a high affinity. This means that safety gloves must be worn at all times to prent the SYBR Green dye from getting on the skin.
A flourimeter uses optical caustics in applied to flourescence to observe and measure the amount of flourescence. Flourescence can occur when a wavelength excites a molecule causing the emission of light. In this experiment, the SYBR Green when attached to the DNA nucleotides should glow when placed on the Flourimeter. Optical caustics causes light to be concentrated near the surface of the drop in order to increase the intensity of the flourescence. This is what can be eventually captured by our cell phones. The flourimeter is made of generic plastic, and a drop is placed on a special type of slides with a rough surface with a micropipette. Once aligned, a blue light is shown through the drop causing the SYBR Green to flouresce and an image to be captured on a cell phone to later be recorded on the computer program ImageJ. How the Fluorescence Technique Works First, the flourimeter is set up by turning on the Blue LED and turning on and putting a smart phone on a cradle in front of the flourimeter. From this position, adjust the flourimeter to the height of the cell phone's camera in order to get a picture of the drop sideways and not from any other angle. This can be done by placing books or other materials underneath the flourimeter. After placing the drop of SYBR Green on the slide and adding the DNA to the dye, the dye will bind to the nucletide in the DNA. So, when the SYBR Green is exposed to the blue light, it will start to flouresce, putting a dark container over the reaction will make the flourescence easier to see in the picture taken by your cell phone.
ProcedureSmart Phone Camera Settings
[Instructions: If you used an additional phone, describe the other type of smart phone you used and how you adjusted the camera settings, if applicable. If not, delete this part]
[Instructions: Describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points.]
[Instructions: See worksheet page 6.]
[Add more rows as needed]
Data AnalysisBasic Set Up of the Flourimeter Procedure Representative Images of Samples
Image J Values for All Samples PCR Product Table
PCR Product Table Calculations
[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.]
Fitting a Straight Line
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