OK, so we all know that Plasmidsaurus nanopore sequencing isn’t perfect. Every time I see the mistake at the 5′ end of the IRES sequence I know to ignore it, but there are bunch of other ones that I still repeatedly run into (but not quite as frequently) such that I don’t have it memorized and am not sure if I should be ignoring it right off the bat. Thus, I’m going to keep a list of repeated erroneous calls here on this page so I’m reminded to ignore them in the future.
Visual evidence of individual example listed below. But here’s a summary of Plasmidsaurus errors to ignore:
- IRES – deletions near the 5′ end
- mCherry – W63R or Q114R
- mScarlet – errors at R71 (sometimes R71G) and S113/L114 (including L114P).
- mKG – L96P
- Puromycin resistance gene PAC – R18G or L125P
- shBleR – Q56R
- Silent or frameshift mutations at the NPGP motif at the 3’end of the P2A sequence
In fact, be very suspicious of any unexpected L -> P mutant through Plasmidsaurus seq. And maybe Q -> R muts too.