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    7/24 PCR Troubleshooting YF1/FixJ
    • Notes
    • Metadata
    • Relevant Items
    • PCR amplification of plasmid DNA (Phusion)
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    7/24 PCR Troubleshooting YF1/FixJ

    Wednesday, 7/24/2019ObjectiveTable1Thursday, 7/25/2019ObjectiveGel 1
    Wednesday, 7/24/2019
    Objective
    We will be testing a different combination of primers and template DNA to gain insight on why PCR of YF1/FixJ has consistently resulted in an incorrect amplicon of length ~1 kb (the correct amplicon should be around 1.8 kb). We will be using fresh template DNA from today (new minipreps of YF1/FixJ).
    What we currently know:
    ●
    The 1 kb band is not a result of reagent contamination given that PCRs of parts other than YF1/FixJ, using the same reagents (water, polymerase, buffer, etc.), have been successful and have not had the 1 kb band
    ●
    The 1 kb band is not a result of contamination of the VR
    Thursday, 7/25/2019
    Objective
    Gel electrophoresis to analyze results of the PCRs above

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