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Publisher Summary Increasing the number of readable nucleotides per sequencing reaction reduces the number of template DNA preparations and sequencing reactions; the sequence assembly from individual reactions can be significantly simplified. DNA sequencing requires high detection sensitivity because a typical band contains about 0.02–1 fmol of DNA. These requirements have been met by radioactive, L2 fluorescent, and enzyme-linked detection systems. Enzyme-linked detection offers advantages for both high throughput and occasional needs. For high throughput, automatic film readers and development automatons can be considerably faster and less costly than fluorescent devices. For occasional sequencing, the higher stability of reagents and sequencing reaction products of nonradioactive systems can be helpful; these advantages can be achieved with significantly lower equipment costs for enzyme-linked detection systems than with fluorescent devices. While the membrane-bound sequence patterns produced by direct transfer electrophoresis can be detected by conventional radioactive labeling, they also offer a starting point for other detection methods that require a membrane-bound pattern, such as genomic sequencing, multiplex sequencing, or sequencing with enzyme-linked detection. |
