Objective: To analyze existence of an association between methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism with male infertility. Materials and methods: A case–control study was conducted from June 2017 to August 2018 in which 88 infertile and 40 fertile were recruited. Polymerase chain reaction (PCR) – restriction fragment length polymorphism (RFLP) assay was carried out to study the allelic frequency of C677T polymorphism. The differences in allelic and genotypic frequencies of C677T locus between fertile and infertile groups were evaluated by the Pearson chisquare test. A logistic regression model was used to calculate Odds ratios and 95% confidence intervals, p value < 0.05 was considered significant. The Hardy–Weinberg equilibrium was tested using HWE software. Results: In infertile subjects, frequency distribution of CC allele was (60.2%), the CT allele was (30.7%) the TT allele was (9.1%) and in the fertile controls the frequency was CC allele (75%), CT allele (20%) and TT allele (5%) respectively. Analysis revealed MTHFR 677 CC genotype associated significantly with male infertility (p < .046, OR = 2.385; 95% CI = 1.014–5.608); Frequency of CT (30.7%) and TT (9.1) genotypes were higher in infertile men as compared to CT (20%) TT (5%) in fertile controls but statistically these were not significantly different (p = 0.097; OR = 0.455; CI = 0.179–1.153 and p = 0.431; OR = 0.526; CI = 0.107–2.599 respectively). Significant association of age and BMI with MTHFR genotypes and infertility was observed. Conclusion: Our results showed that MTHFR C677T polymorphism is not a risk factor for male infertility in our Pakistani population.