Experience in shotgun sequencing a 134 kilobase pair DNA molecule.

Until now, large DNA sequences have been obtained by cloning fragments of the target molecule into plasmid, cosmid or bacteriophage lambda vectors. The 134 kbp DNA sequence of channel catfish virus was determined with relative ease by shotgun cloning of random fragments of genomic DNA directly into a bacteriophage M13 vector, sequencing by dideoxynucleotide chain termination, and compilation of the data using Staden's database handling programs. Experience gained during this endeavour indicates that sequences substantially larger than 134 kbp may be obtained using this approach.