BME103 s2013:T900 Group1: Difference between revisions

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==Protocols==
==Protocols==


'''Thermal Cycler Program'''<br>
'''Thermal Cycler Program'''<br><br>
 
'''Stage 1'''<br>
95°C for 3 minutes: ''Initial DNA strand is separated.''<br>
'''Stage 2'''<br>
35 cycles of the following steps, each with a duration of 30 seconds:<br>
# 95°C: ''Double strands of DNA separate.''<br>
# 57°C: ''Primers attach at ends of target DNA segment.''<br>
# 72°C: ''DNA polymerase activates and replicates target segment of DNA.''<br>
'''Stage 3'''<br>
Final Hold 4°C for 3 minutes: ''PCR reaction is stopped''<br><br>


'''DNA Sample Set-up'''<br>
'''DNA Sample Set-up'''<br>
Two samples of DNA were tested in triplicate. Patient 1's ID number is 29013 and Patient 2's ID number is 13146. The samples for Patient 1 were labeled α1, α2, and α3. Similarly, Patient 2's samples were labeled β1, β2, and β3. Additionally, a positive control (CP) and a negative control (CN) were used. The samples were arranged for PCR reaction as follows:


{| {{table}}
{| {{table}}
|-
|-
| row 1 cell 1 || row 1 cell 2 || row 1 cell 3 || row 1 cell 4
| CP || α1 || α2 || α3
|-
|-
| row 2 cell 1 || row 2 cell 2 || row 2 cell 3 || row 2 cell 4
| CN || β1 || β2 || β3
|}
|}



Revision as of 12:18, 22 March 2013

BME 103 Spring 2013 Home
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Lab Write-Up 1
Lab Write-Up 2
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Kristi Norris
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LAB 1 WRITE-UP

Initial Machine Testing

The Original Design
(Add image of the full OpenPCR machine here, from the Week 9 exercise. Write a paragraph description for visitors who have no idea what this is)


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: Initial DNA strand is separated.
Stage 2
35 cycles of the following steps, each with a duration of 30 seconds:

  1. 95°C: Double strands of DNA separate.
  2. 57°C: Primers attach at ends of target DNA segment.
  3. 72°C: DNA polymerase activates and replicates target segment of DNA.

Stage 3
Final Hold 4°C for 3 minutes: PCR reaction is stopped

DNA Sample Set-up
Two samples of DNA were tested in triplicate. Patient 1's ID number is 29013 and Patient 2's ID number is 13146. The samples for Patient 1 were labeled α1, α2, and α3. Similarly, Patient 2's samples were labeled β1, β2, and β3. Additionally, a positive control (CP) and a negative control (CN) were used. The samples were arranged for PCR reaction as follows:

CP α1 α2 α3
CN β1 β2 β3

DNA Sample Set-up Procedure

  1. Step 1
  2. Step 2
  3. Step 3...

PCR Reaction Mix

  • What is in the PCR reaction mix?

DNA/ primer mix

  • What is in the DNA/ primer mix?






Research and Development

Specific Cancer Marker Detection - The Underlying Technology

(Add a write-up of the information discussed in Week 3's class)

(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 OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)