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    7/20 PCRs
    • Notes
    • Metadata
    • Relevant Items
    • PCR amplification of plasmid DNA (Phusion)
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    7/20 PCRs

    Saturday, 7/20/2019PCRTable2Table7Analytical Gels of PCRsGel17/20 Gel 1Gel 2IMG_20190720_202958.jpg
    Saturday, 7/20/2019
    PCR
    Diluting primers: Primers at 100uM, dilute to 10uM. Make 50uL of it.
    DNA Calculations:
    Diluted by 1:100
    25uL reaction for parts
    15.75uL RNase free water
    Buffer
    1uL template DNA
    20uL reaction for control
    Analytical Gels of PCRs
    7/20 Gel 1
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    weird that positive/negative controls didn't work, but it seems that amilCP was successfully PCRed
    IMG_20190720_202958.jpg
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    YF1 and FixJ are screwed up...entire thing should be 1.8kbp but bands show them at a little less than 1kbp. It's also consistent across 6 trials. We figured VR is good, so our best guess is that the designed primers bound somewhere unexpected in the middle of the sequence. We will be redoing this PCR tomorrow to figure out what went wrong. Apart from that, sfGFP looks good.

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