Sequencing gels, development

Another [3,3]-sigmatropic rearrangement/RCM sequence was developed by Srikrishna and coworkers in the first total synthesis of the aromatic sesquiterpene ( )-laurokamurene B (24) (Scheme 1.5) [7]. RCM of 27 led to cyclopentenol 28 bearing a labile hydroxyl moiety, both benzylic and allylic. The latter compound was found to be unstable and when treated with silica gel delivered the rearranged alcohol 29 in a one-pot operation. 

Eor example, the technique of Southern blotting was developed (68) for use with agarose gel electrophoresis of DNA fragments. Southern blots are designed to detect specific sequences of DNA. After electrophoresis is complete, the DNA is denatured and the single stranded DNA transferred to the specially prepared nitrocellulose paper. The nitrocellulose is then incubated with radioactive RNA or DNA complementary to those DNA sequences of interest. After the nitrocellulose has been sufftciendy incubated with the radioactive complementary DNA, autoradiography is used to identify the fragments of interest. 

The extraordinary complexity of human genes and their products has encouraged the development of extremely high-resolution analytical methods.75 Capillary electrophoresis is competitive with slab gel methods, with resolution up to the order of about 1,000 base pairs for sequencing, sizing, and detection of mutation. Reversed phase HPLC is useful for restriction digest mapping and MALDI-MS up to about 1000 base pairs. 

The use of other crosslinking metals developed simultaneously with the use of antimony, chromium, and boron(borate). Tiner, et al.(242) introduced titanium (IV) crosslinkers in 1975 as ammonium tetralactonate or bis(triethanolamine)bis(isopropyl)titanium(IV). Upon contact with water soluble titanium (IV) derivatives ordinarily form orthotitanic acid, Ti(0H)4, which rapidly forms oligimeric metatitanic acid, [Ti(0H)2] and titanium dioxide. Electron donors such as the hydroxyl groupsxof polysaccharides, if properly oriented, can participate in the sequence of titania reactions and a crosslinked gel network results. Various titanium metal crosslinkers remain in common use today. More will be said about titanium crosslinked gels later. 

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