A total of 507 simple sequence repeats (SSRs or ''microsatellites'') were identified from Plasmodium falciparum sequences in GenBank and from inserts in a genomic DNA library. Oligonucleotide primers from sequences that flank 224 of these SSRs were synthesized and used in PCR assays to test for simple sequence length polymorphisms (SSLPs). Of the 224 SSRs, 188 showed SSLPs among 12 different P. falciparum lines; 116 of these SSLPs were assigned to chromosome Linkage groups by physical mapping and by comparing their inheritance patterns against those of restriction fragment length polymorphism markers in a genetic cross (HE3xDd2). The predominant SSLPs in P. falciparum were found to contain [TA](n), [T](n), and [TAA](n), a feature that-is reminiscent of plant genomes and is consistent with the proposed algal-like origin of malaria parasites. Since such SSLPs are abundant and readily isolated, they are a powerful resource for genetic analysis of P. falciparum. (C) 1996 Academic Press, Inc.