User:Anthony Salvagno/Notebook/Research/2011/01/24/pBSTXI PCR

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Preliminary Work

  • Make thermocycler program.
    • extend for 5 min
    • anneal at 60C
  • dilute some plasmid DNA
    • currently at 100ng/ul
    • make 1.5ng/ul
    • take 5ul and add 333ul to it
  • dilute primers
    • currently at 50uM and 187uM
    • make 10uM each
    • 1:5 and 1:18.7 respectively

Reaction

{{#widget:Google Spreadsheet |key=0Agbdciapt4QZdF84NTd4YVZBLXdWVm5zeVBZeklMTHc |width=500 |height=300 }}

Reaction Program

I created a new program in the thermal cycler. The program is shown in the reaction spreadsheet above, but here are the specifics anyways:

  • program name is PBSTXI
  • it is set to anneal at 60C
  • it is set to extend at 72C for 5 min. This is per the recommendation of 1min per 1kb of DNA, we have 4.5 kb so this should be alright.

Results