I'm doing research on Cacao swollen shoot virus. but right now I'm facing a problem that primers I use can't detect the virus. I have tried using several primers such as those described by Hoffman et al (1997), Kouakou et al (2012), Oro et al (2008), and Quainoo et al (2008). Is there any solution for the problem? thank you
Maybe this can be of help:
Article A Complex of Badnavirus Species Infecting Cacao Reveals Mixe...
Alternatively, I would try the universal degenerate badnavirus primer sets on RT-PCR using DNAse digested total nucleic acids as template.
@Alejandro Olmedo can you provide me the method using degenerate primers on RT-PCR?
Check this article out:
Article Assessment and optimization of rolling circle amplification ...
I would use the primers BadnaFP/BadnaRP, and BadnaMFP/BadnaMRP. I would use a annealing temperature of 45C.
Good luck.
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