Short-insert libraries as a method of problem solving in genome sequencing.

摘要: As the Human Genome Project moves into its sequencing phase, a serious problem has arisen. The same been increasingly vexing in closing phase of Caenorhabditis elegans project. difficulty lies efficiently through certain regions which templates (DNA substrates for process) form complex folded secondary structures that are inaccessible to enzymes. solution, however, is simply break them up. Specifically, offending fragments sonicated heavily and recloned, as much smaller fragments, pUC vector. sequences obtained from resulting library can subsequently be assembled, free effects structure, produce high-quality, complete sequence. Because success simplicity this procedure, we have begun use it all standard primer walking at difficult.