Do I need to have a biological replicate for sequencing Virus - (Apr/09/2014 )

Hi all,

I have one organ per time point from H5N1 infected ducks that is preserved in RNA later. This means that I do not have biological replicates.

Now, If I need to sequence the virus to have the full coding sequence of HA and NA genes of H5N1 to build up a phylogenetic tree, Do I need a biological replicate

-Mohamed 1984-

Conventional sequencing is prone to base miscalls and a few other things that mean that your sequence could be wrong. It would be best if you could get 2 reads for each sequence. Preferably the reads would be in opposite directions.

-bob1-

But is it scientifically wrong for publishing paper contain such an experiment ؟

-Mohamed 1984-

No- think about it this way - if you were sampling wild ducks (are you?) then a different duck could be infected with a different strain of influenza to others on the same pond (maybe it flew in that morning) so how could you have biological replicates?

-bob1-