Apparatus and methods for chemical synthesis
AssigneeAcademia Sinica
InventorsKonan Peck, Ji-Yen Cheng
Life of this patent
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Abstract
Apparatuses and methods are described for parallel oligonucleotide synthesis of hundreds of different sequences and lengths at a time. Standard phosphoramidite chemistry is employed. The syntheses take place in a reaction plate compatible with the industrial standard microplate format to allow the use of readily available automated instruments for subsequent processing. Key parameters in reducing synthesis volume in small reaction wells are discussed. This invention provides solutions to the difficulties of low volume, high number synthetic reactions.
Claims26
An apparatus for chemical synthesis comprising:
- a plate including a plurality of wells, wherein each of the wells defines a bore extending from a first end to a second end of each well, includes a synthesis reaction medium disposed within the well between the first and second ends, and has an opening at the first end having an inner diameter of about 3.5 mm or less, an inner diameter at the synthesis reaction medium closest to the second end of about 3.0 mm or less, and each of the wells are arranged in a 2-dimensional array such that the wells are spaced apart from neighboring wells center-to-center by about 4.5 mm or less;
- and
- a supply assembly including at least one valve for dispensing fluids through a nozzle, wherein the valve and nozzle together are configured to provide fluids to wells in volumes as small as about 2 microliters.
A method of conducting parallel chemical synthesis comprising:
- determining a dispersing pressure for dispensing fluids through a supply assembly into reaction wells of a synthesis apparatus based upon a coefficient of variation for each dispersed fluid as a function of the dispersing pressure and a synthesis support scattering height in the wells for each dispersed fluid as a function of the dispersing pressure.
The method of 13 further comprising dispensing a plurality of reactants via the supply assembly into a plurality of wells of a parallel synthesis plate, wherein each of the wells defines a bore extending from a first end to a second end of each well, includes the synthesis reaction medium disposed within the well between the first and second ends, and has an opening at the first end having an inner diameter of about 3.5 mm or less, an inner diameter at the synthesis reaction medium closest to the second end of about 3.0 mm or less, and each of the wells are arranged in a 2-dimensional array such that the wells are spaced apart from neighboring wells center-to-center by about 4.5 mm or less;
- and the supply assembly includes at least one valve for dispensing fluids through a nozzle, the valve and nozzle together configured to provide fluids to wells in volumes as small as about 2 microliters.
The method of using tightly sealed synthesis retaining medium such as a membrane or a filter frit for containing synthesis supports (e.g CPG) so that no synthesis supports can leak through the medium.
Dependencies are inferred from the claim text.