I am starting an experiment on G6PD mutations where i will be amplifying the 202 and 376 SNPs from human genomic DNA.I would like to know the cycling temperatures being used for the PCR reaction so that I can start my optimization process.Any help, please?
I would advice you to see the primers. if you can't access to softwares as Oligo7, just put the sequence of your primers at one of the tools proposed by the UCSC (http://genome.ucsc.edu/cgi-bin/hgPcr). after the voice of the genome and the assembly, if the PCR is valide, you will have access to the Tm.
send me a mail if needed ([email protected])
fred
Dear Abena
The temperature cycling protocol: 95°C for 1 min, and then 35 cycles of 94° for 1 min, 62°C for 1 min, 72°C for 1 min followed by a final extension at 72°C for 10 min.
Best wishes
Amin
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