biochemistry3096 Posted November 26, 2011 Share Posted November 26, 2011 Hi, I have recently designed a set of primers to amplify a fragment of a gene present in an cDNA template. I have performed a blast search on my primers and the results have shown a hit with my gene of interest. However, after conducing the PCR and cloning my fragment into a vector, i sent it off for sequencing. The results were surprising. Sequence analysis showed that the amplified fragment was from a completely unrelated gene. I was wondering, bearing in mind my designed primers are specific for my fragment of interest, why I amplified a non-specific fragment. Any suggestions would help. Thank you, Biochemistry3096 Link to comment Share on other sites More sharing options...
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