week 10/7 to 10/ 11

 


We talked about how we’d already digested kana with XbaI before and planned to ligate it and do PCR. We also went over how PCR works:

  • PCR is targeted replication. The machine heats up and breaks the hydrogen bonds, so the DNA separates into single strands.

  • The strands keep separating.

  • Primers go in at their kneeling temperature, which for us is 57°C. This tells the polymerase where to start.

  • At 72°C, the polymerase copies the DNA, going from one strand to two.

  • The machine repeats the process for a set time.

  • Also…

    • pdCas9 is 6,700 bps.

    • Deinococcus usually grows between 30°C and 37°C, but Sonora likes 30°C.

    • E. coli grows best at 37°C.

  • ran a gel to check pgK again, but the results didn’t make sense, and we couldn’t figure out what went wrong.

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