OUR TEAM
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LAB 4 WRITE-UP
Protocol
Materials
- Lab Coat and disposable gloves
- PCR reaction mix, 8 tubes, 50 μL each: Mix contains Taq DNA polymerase, MgCl2, and dNTP's (http://www.promega.com/resources/protocols/product-information-sheets/g/gotaq-colorless-master-mix-m714-protocol/)
- DNA/ primer mix, 8 tubes, 50 μL each: Each mix contains a different template DNA. All tubes have the same forward primer and reverse primer.
- A strip of empty PCR tubes
- Disposable pipette tips: only use each only once. Never re-use disposable pipette tips or samples will be cross-contaminated
- Cup for discarded tips
- Micropipettor
- Open PCR machine: shared by two groups
PCR Reaction Sample List
Tube Label |
PCR Reaction Sample |
Patient ID
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G5 + |
Positive control |
none
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G5 - |
Negative control |
none
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G5 1-1 |
Patient 1, replicate 1 |
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G5 1-2 |
Patient 1, replicate 2 |
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G5 1-3 |
Patient 1, replicate 3 |
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G5 2-1 |
Patient 2, replicate 1 |
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G5 2-2 |
Patient 2, replicate 2 |
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G5 2-3 |
Patient 2, replicate 3 |
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DNA Sample Set-up Procedure
- Step 1
- Step 2
- Step 3...
OpenPCR program
- HEATED LID: 100°C
- INITIAL STEP: 95°C for 2 minutes
- NUMBER OF CYCLES: 35
Denature at 95°C for 30 seconds, Anneal at 57°C for 30 seconds, and Extend at 72°C for 30 seconds
- FINAL STEP: 72°C for 2 minutes
Research and Development
PCR - The Underlying Technology
Q1: What is the function of each component of a PCR reaction
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