Restriction mapping of phage λ vectors using non-radioactive methods

In order to take advantage of non-radioactive methods, we have developed two plasmids (pλLE and pλRE) for mapping restriction sites of long inserts cloned in phage λ vectors. These plasmids are constructed by cloning the left 402-bp and right 560-bp phage λ genome ends, respectively. To map restriction sites, the cloned sequences in pλLE and pλRE are labeled with digoxygenin and hybridized to partially digested λ DNA. The ladder of bands detected with these probes can be used to construct restriction maps in the same way as those obtained using radioactively labeled cos complementary oligodeoxyribonucleotides [Rackwitz et al., Gene 30 (1984) 195-200]. © 1994.