BME100 f2013:W1200 Group8 L4: Difference between revisions
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| [[Image: | | [[Image:Scooby.jpg|100px|thumb|Name: Colby Mark<br>Research and Development Specialist]] | ||
| [[Image: | | [[Image:Shaggy.jpg|100px|thumb|Name: Hayden McIver<br>Open PCR Machine Testing]] | ||
| [[Image: | | [[Image:Velma.jpg|100px|thumb|Name: Isai Valdez<br>Protocol Planning]] | ||
| [[Image: | | [[Image:Daphne.jpg|100px|thumb|Name: Madison Grayson<br>Protocol Planning]] | ||
| [[Image: | | [[Image:Fred.jpg|100px|thumb|Name: Andrew Wills<br>Open PCR Machine Testing]] | ||
|} | |} | ||
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'''The Original Design'''<br> | '''The Original Design'''<br> | ||
[[Image:Group8-PM-PCR.JPG]][[Image:PCR5.jpg]] | |||
OpenPCR is a themocycler used to conduct heat through samples of data. This is used to replicate DNA through a series of heating and cooling in order to allow certain chemical reactions to occur.The design is made to prevent any outside reagents from affecting the concentrations of the samples; certain features such as precise temperature control and a heated lid allow there to be no interference with the data. The OpenPCR conveniently connects to your computer and stores the data from every trial. <br> | OpenPCR is a themocycler used to conduct heat through samples of data. This is used to replicate DNA through a series of heating and cooling in order to allow certain chemical reactions to occur.The design is made to prevent any outside reagents from affecting the concentrations of the samples; certain features such as precise temperature control and a heated lid allow there to be no interference with the data. The OpenPCR conveniently connects to your computer and stores the data from every trial. <br> | ||
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'''Experimenting With the Connections'''<br> | '''Experimenting With the Connections'''<br> | ||
When we unplugged (part 3) from | When we unplugged the LCD screen (part 3) from the circuit board (part 6), the machine's LCD screen turned off because power was cut off. | ||
When we unplugged the white wire that connects the circuit board (part 6) to heating block (part 2), the machine showed a drop in temperature because the heating element lost power. | |||
'''Test Run'''<br> | |||
The date that we first tested the Open PCR was on October 23rd, 2013. We didn't think that we would be able to open up the panel and see the inner workings of this machine. We thought it was very interesting to see how the wires are connected and also how the cooling and heating system works with the layered Aluminum plates. The process of completing the entire lab did take quite a long time, but compared to other machines ours worked to perfection. | The date that we first tested the Open PCR was on October 23rd, 2013. We didn't think that we would be able to open up the panel and see the inner workings of this machine. We thought it was very interesting to see how the wires are connected and also how the cooling and heating system works with the layered Aluminum plates. The process of completing the entire lab did take quite a long time, but compared to other machines ours worked to perfection. | ||
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'''Thermal Cycler Program'''<br> | '''Thermal Cycler Program'''<br> | ||
Heated Lid: 100 degrees Celsius<br> | |||
Initial Step: 95 degrees Celsius for 3 mins<br> | |||
Step: 35 cycles<br> | |||
-Denature: 95 degrees C for 30 sec<br> | |||
-Anneal: 57 degrees C for 30 sec<br> | |||
-Extend: 72 degrees C for 30 sec<br> | |||
Final Step: 72 degrees C for 3 mins<br> | |||
Final Hold: 4 degrees C<br> | |||
'''DNA Sample Set-up'''<br> | '''DNA Sample Set-up'''<br> | ||
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{| {{table}} | {| {{table}} | ||
|- | |- | ||
| | | Cancer DNA template (C+) || patient 1 ID:21489 Tube: 1R1 || Patient 1 ID: 24189 Tube: 1R2 ||Patient 1 ID: 24189 Tube: 1R3 | ||
|- | |- | ||
| | | Non-cancer DNA template (C-) || Patient 2 ID:90434 Tube: 2R1 || Patient 2 ID:90434 Tube: 2R2|| Patient 2 ID:90434 Tube: 2R3 | ||
|} | |} | ||
'''DNA Sample Set-up Procedure''' | '''DNA Sample Set-up Procedure''' | ||
# Step 1 | # Step 1. Label each tube to avoid using wrong substance. | ||
# Step 2 | # Step 2. Dispense 50 ul of PCR reaction mix using pipettes into each of the 8 tubes | ||
# Step 3... | # Step 3. Dispense 50 ul of each DNA sample in their respective tubes, switching to a new tip when a different substance is handled to prevent cross contamination. | ||
# Step 4. Place tubes in PCR machine and close the lid. | |||
# Step 5. make sure PCR machine is programmed to the right temperatures and cycles. | |||
# Step 6. Connect the PCR machine to the computer. | |||
# Step 7. Run the Thermo cycler program on the Open PCR software. | |||
'''PCR Reaction Mix''' | '''PCR Reaction Mix''' | ||
* | * The PCR reaction mix contains the following: | ||
-Taw DNA polymerase | |||
-MgCl2 | |||
-dNTP | |||
'''DNA/ primer mix''' | '''DNA/ primer mix''' | ||
* | * -A forward primer | ||
-A reverse Primer | |||
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Latest revision as of 10:44, 30 October 2013
BME 100 Fall 2013 | 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 TEAMLAB 1 WRITE-UPInitial Machine TestingOpenPCR is a themocycler used to conduct heat through samples of data. This is used to replicate DNA through a series of heating and cooling in order to allow certain chemical reactions to occur.The design is made to prevent any outside reagents from affecting the concentrations of the samples; certain features such as precise temperature control and a heated lid allow there to be no interference with the data. The OpenPCR conveniently connects to your computer and stores the data from every trial.
When we unplugged the LCD screen (part 3) from the circuit board (part 6), the machine's LCD screen turned off because power was cut off. When we unplugged the white wire that connects the circuit board (part 6) to heating block (part 2), the machine showed a drop in temperature because the heating element lost power. Test Run
ProtocolsThermal Cycler Program DNA Sample Set-up
-Taw DNA polymerase -MgCl2 -dNTP
-A reverse Primer
Research and DevelopmentPCR - The Underlying Technology
Each step of thermal Cycling Nucleotide Base Pairing
|