This is perhaps more of a philosophical question - I am wondering for Sanger sequencing of a PCR fragment specifically - but is it necessary to do biological replicates for clean Sanger sequencing data?
For example, I recently sequenced an exon of a gene in 2 cell lines - for each of the two cell lines my template DNA came from a pool of 100,000 cells. I PCRed up the fragment and then sequenced it. Is it necessary to repeat this experiment? After all, the template DNA came from many thousands of cells. While the experiment is easy to repeat I'm curious to know whether it's necessary or expected.