In this study we have generated a dose-response curve for the formaldehyde induction of recessive lethal mutations in the eT1(III;V)-balanced region of C. elegans. We have mapped 96 out of 112 formaldehyde-induced lesions to either LGIII or LGV and genetically analyzed 31 lesions that mapped to LGV. Our findings showed that a 4-h treatment with 0.1% formaldehyde gave the best mutation induction frequency with the least side effects. We found that formaldehyde induced putative point mutations, deficiencies and more complex lesions in C. elegans. We isolated 11 putative point mutations, 3 of which defined new genes and 8 were alleles of known genes. One of the new genes, let-450, is currently the left-most known gene on LGV. We also isolated 5 deficiencies. Our formaldehyde-induced lesions increased the number of zones in the eT1-balanced region of LGV from 22 to 34.