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Viral DNA encoding nerve agent model compound-resistant hBChE variants was sequenced to identify sites where mutagenesis was present.
The DNA region containing the 1747delT, 1779T>G, 1808C>G, 1822G>A and 1888insT variants was sequenced in the reverse direction with 0.16 μM primer set B reverse primer and was purified by ethanol precipitation preceded by ExoSAP-IT (USB Corp., Cleveland, OH, USA) clean up, according to the manufacturer's protocol.
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As a control, a fraction of variants were sequenced using the ABI Prism BigDye Terminators and the ABI Prism 3100 sequencer (Applied Biosystems).
Virus variants were sequenced and plasmid-based reverse genetics was used to examine the effect of each mutation.
Over 600 variants were sequenced to compile a comprehensive database of P8 sequence diversity compatible with assembly into the wild-type phage coat.
Noteworthy, the alignment-based approach that was used for the calculation of the total mean sequencing error rate revealed that several MinION reads were composed of non-contiguous regions of the YCNV genome, suggesting that either defective viral variants were sequenced or that the MinION generated chimeric sequences.
The peptides from the H2A variants were sequenced with different PTMs (Table 3, Figure S2A).
The scFv gene fragments of the improved variants were sequenced (figure 1).
Plasmid DNAs extracted from the yeast colonies expressing WT levels of Ynd1 variants were sequenced.
Representatives of all allelic variants were sequenced to ensure the assignment of the number of repeats was correct.
Cloned gene variants were sequenced.
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