Based on the research of predictingβ-hairpin motifs in proteins, we apply Random Forest and Support Vector Machine algorithm to predictβ-hairpin motifs in ArchDB40 dataset. The motifs with the loop length of 2 to 8 amino acid residues are extracted as research object and thefixed-length pattern of 12 amino acids are selected. When using the same characteristic parameters and the same test method, Random Forest algorithm is more effective than Support Vector Machine. In addition, because of Random Forest algorithm doesn’t produce overfitting phenomenon while the dimension of characteristic parameters is higher, we use Random Forest based on higher dimension characteristic parameters to predictβ-hairpin motifs. The better prediction results are obtained; the overall accuracy and Matthew’s correlation coefficient of 5-fold cross-validation achieve 83.3% and 0.59, respectively.