BME100 s2015:Group5 9amL4

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BME 100 Spring 2015 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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OUR TEAM

Name: Jacob Licko
Name: Stephen Belikoff
Name: Khalil Khoury
Name: Daniela Romero
Name: Wei Lu

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
G5 + Positive control none
G5 - Negative control none
G5 1-1 Patient 1, replicate 1
G5 1-2 Patient 1, replicate 2
G5 1-3 Patient 1, replicate 3
G5 2-1 Patient 2, replicate 1
G5 2-2 Patient 2, replicate 2
G5 2-3 Patient 2, replicate 3


DNA Sample Set-up Procedure

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


OpenPCR program

  1. HEATED LID: 100°C
  2. INITIAL STEP: 95°C for 2 minutes
  3. 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

  1. 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