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

WiFi coli: A communicolight system

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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