Random subcloning of sonicated DNA: application to shotgun DNA sequence analysis.
A method for producing random subclones using sonication to fragment the DNA is presented. The sonication is combined with enzymatic repair of the fragment ends and a rigorous size fractionation step to prepare subclones of relatively homogeneous and specific size. Under some conditions sonication is shown to shear A + T-rich sequences preferentially, although under most conditions it will create a random subclone library. The use of these subclone libraries for an improved "shotgun" DNA sequencing strategy is tested on a 17.2-kb (kilobase) fragment of Epstein-Barr virus.
D001483:Base Sequence; D003001:Cloning, Molecular; D004260:DNA Repair; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000302:isolation & purification*; D015246:Deoxyribonuclease EcoRI; D004854:Herpesvirus 4, Human; D013010:Sonication
D004279:DNA, Viral; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI