If the virus is positive single strand ( i.e make as mRNA ) ,I will put the genomic as an input for primer3 then click on right primer and left primer: then it gives us the forward and reverse primers that used for amplification and one of them ( reverse primer ) used for cDNA synthesis to make double strand primer.
but my question is : if the virus single strand negative , do I need to make reverse complement sequence to keep our 5' and 3' ends properly oriented.
by EMBOSS revseq program so the result is sense template (- strand) so I can design primers as previous.
Hi,
I think that making revers complement not necessary because the aim is annealing of primer to temple which is virus genome if you make reverse complement primer may cant attach to its template correctly
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