2.3.4. Segregation analysis in European replication cohort

Filtering was performed for missense and nonsense variants that fulfilled all three of the following criteria: (i) a CADD score ≥15 (Rentzsch et al., 2019); (ii) an MAF ≤0.1% in the GnomAD non‐Finnish European exomes; and (iii) a presence in nsCL/P patients only in the present cohorts (Figure 3b). To determine whether the detected variants encompassed further DNVs, Sanger sequencing was performed in unaffected parents for whom DNA was available. For families with more than one affected family member, co‐segregation analysis using Sanger sequencing was performed when DNA was available. Primer sequences are available upon request. For index patients carrying a DNV, paternity testing was performed using the Powerplex 16 HS System (Promega).