I'm trying to make a standard curve for my Real-time PCR. I need to amplify part of my virus genome which is a ss RNA, positive sense genome. I need to use genomic template by boiling my virus and use its genome as my template, PCR a part of it and ligate it ino a vector from TOPO cloning kit. My question is that whether I need to make a cDNA from the template first and then use it in the PCR reaction or there is another way to perform this process. I've never made such a thing and need some help. Please drop me a line if you have any idea.
Thanks a lot.