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Team:UAB-Barcelona/PCRP
From 2009.igem.org
PCR of phosphate detection sequences
We used GoTaq Green Master Mix (Promega)12
Annealing temperature
phoA (100 pb): as annealing temperature is 74ºC, the temperature we should set in the thermociclator would be 69ºC. But it didn't work properly at this temperature, so we set the temperature to 66ºC and 64ºC. PCR in both cases worked properly
lambda-DNA hinIII; 3 x phoa 64ºC; control without primers; control without DNA template; lambda-DNA hinIII; 3 x phoa 66ºC; control without primers; control without DNA template; lambda-DNA hinIII
Purification of phoA
We purified the bands with the kit Wizard SV Gel and PCR Clean-Up System (PROTOCOL).
Agarose gel phoa purification
From left side to right: lambda-DNA hinIII; 3 x phoa 64ºC; control without primers; control without DNA template; lambda-DNA hinIII; 3 x phoa 66ºC; control without primers; control without DNA template; lambda-DNA hinIII
