4/13 PCR with our Rubisco Primers

Today, the control primers (the Rubisco plant primers) we designed arrived! We used the five test food DNA and the positive MM for 12 reactions (180μL sterilized H2O, 30μL 10x buffer, 6μL DNTPs, and 1.5μL taq). We are going to test each of the five text foods with Dr. Todd’s GMO primers and our Rubisco primers. Before we could use the Rubisco primers, we had to calculate the dilution of primers for the primary stock and working stock.

primer calculations

Dilution of Primers Calculations

We created the 10 PCR reactions below:

New Primers PCR reaction contents

michael pcr reactions with new primers

Michael Making PCR Reactions

We ran the PCR under these conditions:

  • 94 degrees for 2 mins
  • 40 cycles of:
  • 94 degrees for 1 min
  • 59 degrees for 1 min
  • 72 degrees for 2 min
  • 72 degrees for 10 mins
  • Hold at 15 degrees

After PCR was completed, the 10 tubes were held at 4 degrees Celsius in a freezer.

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