Blasting Primers for RT-PCR
Posted 20 January 2010 - 02:28 PM
I designed some pairs of primers to perform RT-PCR on Taqman machine. Whenever I BLAST any pair, there always appear mRNA of my gene of interest as well as "hypothetical proteins" (to both forward and reverse but interestingly, direction is opposite : plus/plus -> plus/minus and plus/minus -> plus/plus. final PCR product is also reasonable size = ~100bp ).
What are the hypothetical proteins? Are they same as my gene? or different? Then, I can not design any primer pair and do RT-PCR!! I could not find primers really specific to my gene.
What can I do?
Thanks in advance.
Posted 20 January 2010 - 04:42 PM
Posted 20 January 2010 - 07:07 PM