Cloning method for DNA fragments using arbitrarily primed PCR

A method of cloning DNA fragments comprising the steps of:

• (a) cleaving a vector DNA of plasmid origin with a restriction enzyme,
• (b) dephosphorylating an end of thus obtained cleaved DNA,
• (c) separately obtaining a mixture of DNA fragments by cleaving a chromosomal DNA of a given organism with a restriction enzyme which creates DNA ends cohesive to the ends crated in step (a) in the vector DNA of plasmid origin,
• (d) obtaining a mixture of ligated DNAs through ligation using the dephosphorylated DNAs obtained in step (b) and the mixture of DNA fragments obtained in step (c),
• (e) amplifying the ligated DNAs by PCR with a vector-specific arbitrary primer for the vector employed in step (a) and one or more non-vector-specific arbitrary primers, using the obtained mixture of ligated DNAs as templates, and at an annealing temperature of not lower than 55° C., and
• (f) introducing into a competent cell a cloning vector into which is incorporated a PCR product thus obtained.