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-   -   cDNA PCR troubleshoot (http://www.molecularstation.com/forum/pcr-polymerase-chain-reaction-forum/69475-cdna-pcr-troubleshoot.html)

bliu94 07-16-2009 09:47 PM

cDNA PCR troubleshoot
 
I'm trying to do PCR using cDNA (confirmed to be of good quality w/ nanodrop and PCR with control primers - GAPDH). The primers I am trying this with are for the gene MMP14 and were designed using the NCBI primer designing tool. I inputted the refseq for the mRNA and used the primers it returned. when I do the PCR there are not even smears - only a collection of primers at the bottom. I've tried annealing temps from 50 to 60 degrees and nothing has worked. The product is 600 bp and i'm using taq hotstart. any advice would be appreciated!


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