| 摘要 |
Preparative synthesis of polynucleotides (I) using a microfluidics system (MS), is new. A starter oligonucleotide (ON) is either bound (directly or through a spacer) to the surface of the reaction space in MS, or is synthesized directly, with one end bound to the surface. ON corresponds to one end of (I) being synthesized. One or more rounds of hybridization of 'sequemers', each corresponding to a segment of (I) or its complement and having overlapping ends that allow hybridization, is performed. If required blunt ends, formed e.g. after a polymerase reaction, are converted to sticky ends suitable for hybridization. Where there are nicks between consecutive sequemers, ligation is performed, or if a segment of one or more nucleotides (nt) is missing (complementary to a segment of sequemer) this is completed by polymerase reaction, and ligation of the resulting nick if necessary. The process is repeated as necessary to assemble (I). Independent claims are also included for the following: (1) microfluidics element that includes bound polynucleotides produced by the new method, or their precursors; and (2) a kit or apparatus for the new method comprising appropriately coated microfluidics elements and optionally other components.
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