SYBR Green Dye The process of the lab was to use the green dye to test if the green dye and the DNA shows color when placed in the fluorimeter device. The first step was to place 80uL drop of SYBR Green Dye I solution on the first 2 clear circles in the middle of the slide. The second step was to place 80uL of sample calibration solution on top of the SYBR Green dye. The next step was to adjust the slide so that the blue light can go through the center of the drop of the SYBR Green Dye.
Single-Drop Fluorimeter The fluorimeter device acts as the main part of this lab and it is very vital. The flourimeter acts as the base to where it illuminates blue light through the solution. The fluorimeter heavily depends on the smart phone to capture the solution and the light that goes through it. The flourimeter basically measures the amount of light that illuminates through the SYBR Green Dye and the DNA
How the Fluorescence Technique Works The technique was to use the steps that were given and repeat it until there is five possible measurement position on the slides. The lab was required to add sample calibration of solution on top of the SYBR Green Dye at least five times to gather data.
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:
Flash:
ISO setting:
White Balance:
Exposure:
Saturation:
Contrast:
[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]
Type of Smartphone:
Flash:
ISO setting:
White Balance:
Exposure:
Saturation:
Contrast:
Calibration
First make sure the fluorimeter is on a flat surface. Then use the cradle to hold your smartphone in place. The camera must be at a right angle from the slide in order to take a picture of the drop sideways. Adjust the height of the fluorimeter by turning the nob holding the plastic tray in place. This will help the camera take a better angle of the drop. The distance between the smartphone and the fluorimeter is then measured with a regular ruler. This distance is held constant in order to get a constant yet accurate image of each individual drop. If the distances are different for each trial then there will be a significant difference from one image to the next due to differences in the light collected. The smartphone should be at least 4cm away from the drop. If not, adjust the smartphone so that the smartphone with its cradle is as close to the first two rows of the slide without taking a blurry image. Once the smartphone is adjusted, take a standard ruler and measure the distance between the smartphone and fluorimeter. It is important not to move the camera, cradle, or fluorimeter, because this is the official position that the smartphone needs to be at throughout the whole experiment.
Distance between the smart phone cradle and drop = 5cm
Solutions Used for Calibration
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
Image of no DNA in SYBR Green I
Image of 5 microg/mL of DNA in SYBR Green I
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]
BME 100 Fall 2013
Placing Samples onto the Fluorimeter
