BME100 f2013:W1200 Group13 L4: Difference between revisions

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'''DNA Sample Set-up Procedure'''
'''DNA Sample Set-up Procedure'''
# Step 1
# Step 1 Place each smaple in its respective pipette tips. That is the positive control, negative control, patient 1, and patient 2.
# Step 2
# Step 2 Place the 4 pipettes inside the OpenPCR machine.
# Step 3...
# Step 3 Run the machine for 1 cycle and record results.
# Step 4 Repeat step 3 untill the desired number of replications is reached.




'''PCR Reaction Mix'''
'''PCR Reaction Mix'''
* What is in the PCR reaction mix?
* What is in the PCR reaction mix?
 
The PCR reaction , 50uL each. the mix contains Taq DNA polymerase, MGCl2, and dNTP's.


'''DNA/ primer mix'''
'''DNA/ primer mix'''
* What is in the DNA/ primer mix?
* What is in the DNA/ primer mix?
 
Each mix contains a differant template DNA all tubes have the same forward primer and reverse primer.





Revision as of 13:37, 23 October 2013

BME 100 Fall 2013 Home
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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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LAB 1 WRITE-UP

Initial Machine Testing

The Original Design

The picture above shows what the OpenPCR machine looks like while running and connected to the computer. The machine case is made of wood panels which contains the actual hardware of the device. The smaller box located on the top is the lid of the OpenPCR machine. This contains a 16-tube PCR block which is where the test vials are placed in order to complete the test run. The top part of the lid has a heat plate that heats the vials and also seals the experiment. Inside of the main body of the OpenPCR machine there is a heatsink, processor, led screen, and copper wire connections. The processor has a circuit board that recieves the data from and computes the calculations. The heatsink dispersses the heat and the fan attached cools down the device in order to avoid overheating. The led screen displays the data specifically the numbers of tests run and temperature. The machine also has an USB port and power connection in order to power the device and connect it to the computer in order to run the tests.

Experimenting With the Connections

When we unplugged (part 3) from (part 6), the machine ... (did what? fill in your answer)

When we unplugged the white wire that connects (part 6) to (part 2), the machine ... (did what? fill in your answer)


Test Run

(Write the date you first tested Open PCR and your experience(s) with the machine)




Protocols

Thermal Cycler Program
Stage 1: 95°C for 3 minutes-Double helix unwinds into one long strand
Stage 2: (35 cycles of each)
1. 95°C for 30 seconds-Double helix breaks apart into two strands
2. 52°C for 30 seconds-Primers attach to target sequence
3. 72°C for 30 seconds-DNA polymerase begins constructing new strand
Stage 3: 4°C for 3 minutes-Keeps solution of pure taget sequence at stable temperature

DNA Sample Set-up

PC 1R1 1R2 1R3
NC 2R1 2R2 2R3

DNA Sample Set-up Procedure

  1. Step 1 Place each smaple in its respective pipette tips. That is the positive control, negative control, patient 1, and patient 2.
  2. Step 2 Place the 4 pipettes inside the OpenPCR machine.
  3. Step 3 Run the machine for 1 cycle and record results.
  4. Step 4 Repeat step 3 untill the desired number of replications is reached.


PCR Reaction Mix

  • What is in the PCR reaction mix?

The PCR reaction , 50uL each. the mix contains Taq DNA polymerase, MGCl2, and dNTP's.

DNA/ primer mix

  • What is in the DNA/ primer mix?

Each mix contains a differant template DNA all tubes have the same forward primer and reverse primer.




Research and Development

PCR - The Underlying Technology

(Add a write-up, essay-style, organized into paragrpahs with descriptive headers, based on the Q&A's from Section three of your worksheet)

(BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to credit the sources if you borrow images.)