Improved methods of amplifying DNA in vitro, and genomic cloning and mapping

摘要

A method for cloning genomic DNA comprises the steps of: (a) digesting genomic DNA with a restriction enzyme that generates multiple different cohesive-end sequences; (b) contacting the digested genomic DNA with a linker, said linker comprising a reporter nucleotide sequence and a cohesive-end sequence complementary to a cohesive-end sequence generated by the restriction enzyme of step (a) under conditions such that a linker is joined to a restriction fragment of genomic DNA; and (c) amplifying the product of step (b). Likewise, a method for mapping genomic DNA restriction fragments comprises the steps of: (a) digesting genomic DNA with a restriction enzyme that generates multiple different cohesive-end sequences; (b) amplifying the restriction fragment products of step (a); (c) determining the cohesive-end sequence of each of said restriction fragments; and in accordance therewith, (d) aligning the restriction fragments in a physical map.