sequencing deep mutational scanning library
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12 weeks ago
reaset41 • 0

I would like to do a deep mutational scan of a 1kb bacterial protein. I plan to randomly introduce SNPs with error prone PCR. Then, I will apply a selection filter to evaluate fitness of the variants. My question is regarding sequencing of the resulting pools.

Is it possible to evaluate SNPs from short-read sequencing? (I am worried about the assembly of very similar reads). Or should I introduce a barcode to the variants, link barcodes and SNPs via long-read sequencing, and then sequence barcodes via short-read sequencing to determine the selection?

Thanks!

mutagenesis illumina long-read sequencing • 211 views
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