BME103:W930 Group9 l2: Difference between revisions
Seth Howell (talk | contribs) |
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-100 mL cyber green<br> | -100 mL cyber green<br> | ||
-15 5 piece test tubes<br> | -15 5 piece test tubes<br> | ||
- I quart of DNA mixture including the following per 100 microliters( 10 micro liters forward primer and reverse primer, 50 micro liters of GoTag master mix and 47.8 micro liters of distliled water.) <br><br> | - I quart of DNA priming mixture including the following per 100 microliters( 10 micro liters forward primer and reverse primer, 50 micro liters of GoTag master mix and 47.8 micro liters of distliled water.) <br><br> | ||
'''Supplied by the User'''<br> | '''Supplied by the User'''<br> | ||
-Pipets(100)<br> | -Pipets(100)<br> | ||
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-Micro pipets to add the DNAv | -Micro pipets to add the DNAv | ||
-.2 microliters DNA<br> | -.2 microliters DNA<br> | ||
- | -image J software | ||
<!--- Place your two tables "Supplied in the kit" and "Supplied by User" here ---> | <!--- Place your two tables "Supplied in the kit" and "Supplied by User" here ---> | ||
'''PCR Protocol''' | '''PCR Protocol'''<br> | ||
1. Plug PCR machine in and attach the keypad to USB. <br> | |||
2. Program the PCR heating procedure:<br> | |||
a. Enter in the number of parts the heating portion will have(example First cyle, main cycle and last cycle=3).<br> | |||
b. Enter the number of cycles for each part in order<br> | |||
c. Enter temperature of each cycle in order. <br> | |||
d. place all the test tubes prepared in steps 3-5 in heating pad<br> | |||
e. press * to begin the heating program and wait until the process is complete. <br> | |||
3. Put .2 micro liters of DNA in each test tube. <br> | |||
4. Use a pipet and transfer the 98.8 micro liters DNA priming mixture to each test tube .2 micro liters of DNA.<br> | |||
a. Use one pipet for each transfer do not reuse the pipet.<br> | |||
5. After the mixture is transferred move the tube into the PCR machine.<br> | |||
Revision as of 11:19, 28 November 2012
BME 103 Fall 2012 | Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help | ||||||
OUR TEAMEveryone has contributed to this project even though there are only two usernames. Every person used these two users to make edits to the wiki. Dr. Haynes said that this would be sufficient enough to give each member full participation credit for this project LAB 2 WRITE-UPThermal Cycler EngineeringOur re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski.
The KeyPad will be detachable, and will be connected through the USB connection. Key Features
The key features of the new design include a larger main heating block and a Instructions The new OpenPCR will be assembled and operated almost identical to the old OpenPCR.
ProtocolsMaterials
PCR Protocol a. Enter in the number of parts the heating portion will have(example First cyle, main cycle and last cycle=3). 3. Put .2 micro liters of DNA in each test tube. a. Use one pipet for each transfer do not reuse the pipet. 5. After the mixture is transferred move the tube into the PCR machine.
Flourimeter 1. Place the flourimeter on the table and turn on the blue light. 2. Place provided glass slide on flourimeter track so that the first row of dots is even with the light. 3. Place phone cradle in front of the flourimeter with a smart phone facing perpendicular to the beam of light. (as seen in the picture). 4. Add two drops of green dye on the dots that are even with the light. 5. Place two drops of DNA sample on top of the green dye. 6. Cover the flourimeter and phone by turning the large box over and placing it above both of them. 7. Take a picture of the droplet with the camera. 8. Save the picture and send it to the imageJ operator. Instructions for opening images in imageJ 1. Take a picture of the fluorimeter assembly with a smartphone. 2. Transfer the picture to a laptop equipped with imageJ via icloud or email. 3. Open imageJ and select file, then open. 4. Find the file on the computer and select it. 5. The image is now open and can be analyzed. 6. The image can be split into three images (blue, green, and red) for better analysis by selecting: image-color-split channels. Research and DevelopmentBackground on Disease Markers
Primer Design
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