| 摘要 |
Methods are disclosed for producing a sequence tag from a polynucleotide. In preferred embodiments, the method comprises (a) digesting the polynucleotide with a type II restriction enzyme to produce a first cohesive end; (b) providing a promoter-linker cassette comprising an RNA polymerase promoter, a restriction site for a type IIS restriction enzyme and a second cohesive end which is complementary to the first cohesive end produced by the type II restriction enzyme; (c) ligating the digested polynucleotide to the cassette by joining the first cohesive end and the second cohesive end; (d) digesting the ligated product of step (c) with the type Iis restriction enzyme; and (e) transcribing the resulting construct from the promoter thereby producing a sequence tagged polyribonucleotide.
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