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US 2003/0228602 A1Published application
Solid phase methods for polynucleotide production
AssigneeBlue Heron Biotechnology, Inc.
InventorsHsing-Yeh Parker, John T. Mulligan
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AssigneeBlue Heron Biotechnology, Inc.
InventorsHsing-Yeh Parker, John T. Mulligan
Estimating the term…
Polynucleotides having in excess of 1,000 nucleotides can be prepared using a solid phase synthesis technique. A feature of the technique is the use of a reusable solid support that contains covalently bound oligonucleotide. This covalently bound oligonucleotide is annealed to a bridge oligonucleotide, where the bridge is also annealed to a first oligonucleotide that forms a portion of the target polynucleotide. After the target polynucleotide is synthesized, it can be removed from the solid support under denaturing conditions, and the solid support re-used to prepare additional target polynucleotides. The yield of the target polynucleotide increases when shearing force is applied to the solid support that is linked to the growing oligonucleotide. This shearing force is thought to extend the growing end of the oligonucleotide away from contact with other oligonucleotide bound to the solid support and make that end more accessible to annealing with solution oligonucleotide. The synthesis is conveniently accomplished on a porous frit, where reagents and washing solutions are pumped through the frit.
A method for gene assembly, comprising:
A composition comprising:
An article comprising a solid support coupled to a universal oligo, where one or more of a phosphate group and a polyoxyalkylene group is located between the solid support and a terminal nucleotide of the universal oligo.
37 The article of claim 29 further comprising a bridge oligo, where the bridge oligo comprises a linker polynucleotide region that is annealed to five or more nucleotides of the universal oligo.
In a method for polynucleotide assembly on a solid support in an aqueous environment, the improvement comprising covalently coupling a universal oligo to a solid support either directly or through a linker group, annealing a bridge oligo to the universal oligo to form a starting duplex, the starting duplex having a portion of the bridge oligo in single stranded form to provide a sticky end, and hybridizing a first oligo or a first duplex to the sticky end of the starting duplex, where the first oligo or first duplex is subsequently subjected to ligation conditions and becomes incorporated into a target polynucleotide or fragment thereof.
A method for assembling a portion of a gene on a solid support, the method comprising:
A method for assembling a portion of a gene in solution, the method comprising:
A method for gene assembly, comprising
A method for gene assembly, comprising
A device for automated gene assembly, comprising
Dependencies are inferred from the claim text.