Overview
Shotgun sequencing of a large segment of DNA involves random fragmentation of the target region into smaller segments that are subsequently cloned into a bacteriophage M13 vector. The goal is to create a library of overlapping clones that provide at least fivefold coverage over the entire length of the target fragment. In this protocol, the target segment is sheared by sonication or nebulization. Various sonication methods are described in Table 12-1 on page 12.11 of the print version of the manual. The termini of the resulting DNA fragments are repaired, phosphorylated, and fractionated according to size by gel electrophoresis.
Joseph Sambrook and David W. Russell
Details
Details of this protocol, Generation Of A Library Of Randomly Overlapping DNA Inserts (Subscription Required), are located on a web site other than Biocompare Protocols.