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The amplification program was carried out on a CFX96 instrument, and primers were designed on the based on NCBI data ( Table 1 )

The desalted sample was evaporated to dryness in a speedvac and then resuspended in 10 L Buffer A and analyzed by mass-spectrometry using the following LC-MS gradient: 5% buffer B in buffer A from 0-15 min, 515% buffer B from 1517.5 min, 1535% buffer B from 17.592.5 min, 3595% buffer B from 92.595 min, 95% buffer B from 95105 min, 955% buffer B from 105107 min, and 5% buffer B from 107125 min

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Notably, lack of replication in publicly available datasets does not necessarily exclude biological relevance, particularly for variants that may be ancestry-specific or underpowered in prior studies

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