The cDNA clone T1B was used to detect a 3-allele TaqI restriction fragment length polymorphism (RFLP) in the human topoisomerase I pseudogene locus on chromosome 22q11.2-13.1.Band sizes observed were 8.4 kb (A1), 7.5 Kb (A2), and 5.2 Kb (A3) whose resp. frequencies were 0.34, 0.59, and 0.07 in 70 unrelated caucasians.Constant bands were observed at 6.5, 4.9, 3.9, 3.3, 2.8, 2.1, 1.6, 1.4, and at 1.0 kb.The 2.1-kb constant band mapped to chromosome 1q, whereas all other constant bands mapped to chromosome 20 which contains the active gene for topoisomerase 1.Co-dominant segregation of the RFLP was demonstrated in a 3-generation family.