UMI for Exome Target Sequencing

Question

[alessandro.pastore]

Hi I have a question regarding the use of UMI for Target Re-sequencing.

Incorporating and UMI with the use of a PCR / Amplicon base methods is quite clear for me. But I would like to know How is possible to incorporate an UMI with a target pull down such an Exome or smaller bait set.

Thanks!

[Carlos Borroto]

I'm currently looking for how to do phasing with targeted sequencing data. I wonder if that's the reason you are looking into adding UMI to an exome analysis. I'll be highly interested in seeing if there are any valid answers to your question.

Answer 1

[WouterDeCoster]

After (and perhaps before) the pulldown you normally do a PCR amplification, wouldn't it be possible in your assay to incorporate random nucleotides there in the primer(s) used for amplification? Could you be a bit more specific on your library prep?