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Do I need to have a biological replicate for sequencing Virus


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#1 Mohamed 1984

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Posted 09 April 2014 - 10:34 AM

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  



#2 bob1

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Posted 09 April 2014 - 12:50 PM

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.



#3 Mohamed 1984

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Posted 09 April 2014 - 02:34 PM

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



#4 bob1

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Posted 09 April 2014 - 02:52 PM

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?






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