Dispensable chromosomes in addition to the normal complement in diverse taxa are called B chromosomes. The maize B chromosome is discernible in mitotic chromosome spreads as a small compact chromosome composed mainly of heterochromatin. Although much of this chromosome consists of repetitive elements common to the A chromosomes, several sequences specific to the B chromosome have been identified. In the work described here we used the sequence from a B-specific RAPD (random amplification of polymorphic DNA) marker, pBGBM18.2, to isolate another DNA element, StarkB, present in many copies on the B chromosome. StarkB was mapped to the third and fourth blocks of distal heterochromatin using translocation breakpoints and fluorescent in-situ hybridization (FISH). Sequence analysis revealed that StarkB is composed of fragments from the A genome as well as B-specific sequences. The StarkB element is much larger than the other B-specific elements and is not present in large tandem arrays. Different copies of StarkB varied by small insertions, deletions, and duplications as well as single-nucleotide polymorphisms. Reverse transcriptase PCR showed that portions of the StarkB element are expressed. Using the LTR divergence of retroelements interrupting the B-specific sequences, the minimum age of the StarkB repeat array and, by inference, of the B chromosome, was estimated to be 2 million years. © 2007 Springer.