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Plasmids for producing membrane-displayed somatostatin without FLAG tag (Table 2) were constructed as follows.
All other subunits in the recombinant complex, as well as the synthetic U1 snRNA, were full-length as in the wild type complex; the only exceptions being U1-C which was not present, U1-70k whicomprisedsed residues 1 216 and Sm-G* which contained a C-terminal tag (table S9).
Three primers were designed for each tag (Table 1).
From this set, reads that could be aligned to the Stowers' repeat tag table were excluded (the repeat tag table contains any reads aligned to ≥ 2 locations, unless all locations are from the same gene).
Over 40% sequences in the reference tag database could be mapped to a unique tag (Table 3).
The observed allele size of all individuals genotyped was reduced by 18 bp to account for the M13(–21) universal sequence tag (Table 1).
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Only reads that perfectly matched to tag tables without mis-matches and gaps were considered.
On average 54% of the reads aligned to canonical, 6% aligned to mitochondrial, and <2% aligned to rRNA tag tables.
The remaining reads aligned to repeat (~7%), non-canonical (~3%), non-coding (~0.02%), and intergenic (~14%) tag tables.
For simplicities of comparisons to MAQC results, we merged the canonical, mitochondrial, and rRNA tag tables into one combined table in this manuscript.
In general, the size of the fish that were tagged (Table 1), untagged (refer to Methods section) and sampled for physiological parameters (Tables S2, S3, S4) were approximately equal.
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