Mapping by Multiple Endonuclease Digestions
ثبت نشده
چکیده
Construction of an accurate map of the sites where restriction endonucleases cleave DNA—restriction mapping—is critical for almost all subsequent manipulations of genetic material. Restriction mapping is based upon the cleavage of DNA at or near specific nucleotide sequences with restriction endonucleases and the determination of the sizes of the resulting DNA fragments by gel electrophoresis (see UNITS 2.5 and 2.7).
منابع مشابه
Similar DNA restriction endonuclease profiles in strains of Legionella pneumophila from different serogroups.
DNA of strains of Legionella pneumophila serogroups 1, 3, 4, and 6, isolated from patients and environmental sources, was examined by restriction endonuclease analysis (REA). Major differences in profiles enabled subtyping in many strains with the same serogroup antigen. However, a cluster of L. pneumophila strains, originating from all the examined serogroups, had similar restriction endonucle...
متن کاملPlasmid mapping computer program
Three new computer algorithms are described which rapidly order the restriction fragments of a plasmid DNA which has been cleaved with two restriction endonucleases in single and double digestions. Two of the algorithms are contained within a single computer program (called MPCIRC). The Rule-Oriented algorithm, constructs all logical circular map solutions within sixty seconds (14 double-digest...
متن کاملMolecular cloning and physical mapping of varicella-zoster virus DNA.
Varicella-zoster virus (VZV) DNA was cleaved with restriction endonuclease EcoRI, and most of the resulting fragments were successfully cloned in the phage vector lambda gtWES . lambda B. Double digestions of cloned fragments with EcoRI and BamHI and hybridizations to blot-transferred BamHI digests of VZV DNA were used to construct a physical map of the genome. The molecular termini of the DNA ...
متن کاملApplication of oligonucleotide activation to restriction endonuclease NarI.
Restriction endonuclease NarI cleaves DNA using a two-site mechanism, placing it in the Type IIe class of restriction endonucleases. Although these enzymes have very useful recognition sequences, the two-site mechanism limits the practical application. Site preferences often cause incomplete substrate digestion. Oligonucleotide activation of NarI eliminates incomplete digestions, making it poss...
متن کاملDNA analysis of insect iridescent virus 6: evidence for circular permutation and terminal redundancy.
DNA analysis of small insect iridovirus 6 was performed. Combined exonuclease-restriction endonuclease digestions revealed that all resulting fragments were degraded without preference for any one DNA fragment. Upon denaturation and reannealing of native linear Chilo iridescent virus DNA (158 x 10(6) daltons), duplex DNA circles of a smaller size (140 x 10(6) daltons) with protruding tails were...
متن کامل