Geosmin and 2-methylisoborneol (MIB) are muddy/earthy off-flavor metabolites produced by a range of bacteria. Cyanobacteria are the major producers of the volatile metabolites geosmin and MIB which produce taste and odor problems in drinking water and fish worldwide. Here we detected geosmin and MIB by studying 100 cyanobacteria strains using solid phase microextraction gas chromatography mass spectrometry (SPME GC-MS). A total of 21 geosmin producers were identified from six cyanobacteria genera. Two of the geosmin producers also produced MIB. A PCR protocol for the detection of geoA and MIB synthase genes involved in the biosynthesis of geosmin and MIB was developed. The geoA and MIB synthase genes were detected in all strains shown to produce geosmin and MIB, respectively. Cyanobacterial geoA and MIB synthase sequences showed homology to terpene synthases genes of actinobacteria and proteobacteria. Additional off-flavor compounds, nor-carotenoids β-ionone and β-cyclocitral, were found from 55 strains among the 100 cyanobacterial strains studied; β-ionone was present in 45 and β-cyclocitral in 10 strains. Six of the cyanobacteria which contain off-flavor compounds also produced toxins, anatoxin-a or microcystins. The molecular method developed is a useful tool in monitoring potential cyanobacterial producers of geosmin and MIB.