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Hello community,
I currently have a designed custom amplicon panel with insert sizes around 100bp which for the most part work fine. I'm noticing some variability in that the longer insert sizes seem to have a consistently higher coverage than the shorter ones (lower vs upper quartile). Unfortunately qPCR and dPCR are not options due to the size of the panel.
Platform: MiSeq
Kit/Adapters: TruSeq
Before I redesign my primers, any advice? Is it possible to reliably reduce the insert size to ~50bp or lower and if so, what complications or design considerations are there? I've already lengthened the primer+adapters to somewhat already diminished fragment size.
I currently have a designed custom amplicon panel with insert sizes around 100bp which for the most part work fine. I'm noticing some variability in that the longer insert sizes seem to have a consistently higher coverage than the shorter ones (lower vs upper quartile). Unfortunately qPCR and dPCR are not options due to the size of the panel.
Platform: MiSeq
Kit/Adapters: TruSeq
Before I redesign my primers, any advice? Is it possible to reliably reduce the insert size to ~50bp or lower and if so, what complications or design considerations are there? I've already lengthened the primer+adapters to somewhat already diminished fragment size.