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.