BACKGROUND: Cleft lip with or without cleft palate (CLP) and cleft palate only (CPO) have an inherited component and, many studies suggest, a relationship with folate. Attempts to find folate-related genes associated with clefts have, however, often been inconclusive. This study examined four SNPS related to folate metabolism (MTHFR 677 C -> T, MTHFR 1298 A -> C, MTHFD1 1958 G -> A, and TC II 776 C -> G) in a large Irish population to clarify their relationship with clefts. METHODS: Cases and their parents were recruited from major surgical centers performing cleft repairs in Ireland and a support organization. Data on risk factors, medical history, and DNA were collected. Controls were pregnant women from the greater Dublin area (n = 1,599). RESULTS: CLP cases numbered 536 and CPO cases 426 after exclusions. CPO mothers were significantly more likely than controls to be MTHFR 677 TT, OR 1.50 (95% CI: 1.05-2.16; p = .03). Log-linear analysis showed a borderline association (p =.07). Isolated CPO case mothers were significantly more likely than controls to be homozygous for the MTHFD1 1958 G -> A. variant, OR 1.50 (95%CI: 1.08-2.09; p = .02). When multiple cases were added, both CPO cases and cast, mothers were significantly more likely to be AA (p = .02 and p = .007, respectively). The CLP case-control and mother-control analyses also showed significant effects, ORs 1.38 (95% CI: 1.05-1.82; p =.03) and 1.39 (95%, CI: 1.04-1.85; p =.03), respectively. CONCLUSIONS: Associations were found for both CPO and CLP and MTHFD1 1958 G -> A in cases and case mothers. MTHFR 677 C -> T could be a maternal risk factor for clefts but the association was not strong. Because multiple comparisons were made, these findings require additional investigation. Given the known association between MTHFD1 1958 G -> A and NTDs, these findings should be explored in more detail.