i’ve got a result from my sequencing. when i blast it (eg. aattgcg), it is not related to the spesies that i wan. when i blast it reversely (eg. gcgttaa), it shows that it is related to my spesies. how can we explain it?
Or sequence may be in that manner…….? I guess…
if it is a an inversion mutation, how can the primers bind the sequence correct during RT-PCR as the 5′ and 3′ is already diff? thanks