User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/05/24

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PCR: Second plan

  • Today, Miguel helped me to do the PCR (30 cycles) in order to amplify the luciferase (as band purification failed in two previous occasions).

  • The reaction was done as follows for a total of 30μl:

-H2O ---------------------------------> 13.5μl
-Buffer -------------------------------> 6μl
-Mg(OAc) ----------------------------> 3μl
-dNTPs(0.4mM c/u) ----------------> 2μl
-oligo FWD (prefix, 5pmol/μL) ---> 2μl
-oligo RVS (suffix, 5pmol/μL) ----> 2μl
-DNA ---------------------------------> 1μl
-rtTh(high fidelity)-----------------> 0.5μl

  • The controls were:

-positive: plasmid 18
-negative: blank