LAB 2 WRITE-UP
Thermal Cycler Engineering
Our re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski.
Supplied in the Kit ---- Amount
10 μM Reverse Primer---7.5μL
10 μM Forward Primer---7.5μL
GoTaq master mix--------360μL
Supplied by User-----Amount
Sample of DNA----------.5μL
Computer/Program to run PCR/Image J
DNA Measurement Protocol
1. Remove fluorometer elements from container.
2. Place smartphone into the black stand, creating the camera setup.
3. Position camera setup parallel to the LED box so that the camera is shooting along the channel of the LED box.
4. Place Fluorometer Setup Part I onto lid from original container
5. While leaving the camera setup side of the container unsnapped, place container around Fluorometer Setup Part I.
1. Remove container covering Fluorometer Final Setup.
2. Switch on blue LED.
3. Change camera settings by turning flash on, setting ISO to 800+, increasing exposure to maximum and disabling auto-focus.
4. Pipette two drops of water (volume of 130-160 microliters per drop) onto the slide in the channel of the LED box.
5. Focus LED light on the drops so that the middle of the drop is aligned with the line of light created by the LED.
6. Put the container covering back over the Fluorometer Final Setup.
6. Take 3 photos, making sure not to disturb the positioning and setup.
7. Remove the container covering while again taking care not to change the placement of the setup.
8. Remove slide and dispose of the used slide.
9. Place new slide into LED box, and do steps 4 through 6 again.
10. Note the type of smartphone used throughout the protocol, the distance from smartphone to the base of the LED box (in centimeters), and attach an image of each position of the drops.
Transferal of Fluorometer Data to ImageJ
1. Use a USB cable to connect the smartphone that was used to a computer with the ImageJ software.
2. Open the Start Menu and double click on My Computer or Computer.
3. Select Portable Devices where the smartphone should be listed and double-click the icon.
4. Locate the DCIM folder and open it, and continue to the sub-folder Camera.
5. Select each picture that needs to be transferred, then Copy and Paste them into the folder you choose to store them in.
6. Open ImageJ software from Start Menu or by double-clicking the ImageJ icon on the Desktop.
7. Once ImageJ software is open, click on File and select Open in the toolbar located in the top left of the computer screen.
8. Select Browse then select the image you want out of the folder where you stored your photos (in step 5)
9. Repeat steps 7 and 8 to continue process with different pictures.
Research and Development
Background on Disease Markers