|Taking a directed approach. One directed sequencing strategy exploits a naturally occurring genetic element known as a transposon. The starting point is an ordered set of subclones, each about 3000 base pairs long, derived from a much larger clone (say, a YAC). For each subclone, a preparation is then made in which transposons insert themselves randomly into the subclone -- on average, one transposon in each 3000-base-pair strand. The positions of the transposons are mapped, and a set of strands is selected such that the insertion points are about 300 base pairs apart. Sequencing then proceeds in both directions from the transposon insertion points, using the known transposon sequence as a primer. The full set of overlapping regions yields the sequence for the entire subclone, and the sequences of the full set of subclones yield the sequence for the larger original clone.|
To Know Ourselveswas prepared at the request of the U.S. Department of Energy, Office of Health and Environmental Research, as an overview of the Human Genome Project.