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Porous resin bead and production method of nucleic acid by using same

A porous resin and monomer technology, applied in chemical instruments and methods, preparation of sugar derivatives, library creation, etc., can solve the problems of reducing synthetic nucleic acid, unusable, reducing production efficiency, etc., and achieve the effect of increasing yield

Active Publication Date: 2015-06-10
NITTO DENKO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the property of low-crosslinked and highly swollen resin beads to which many linkers can be bound as origins of nucleic acid synthesis cannot be utilized, and the number of resin beads that can be filled in a column for nucleic acid synthesis decreases, This in turn reduces the amount of synthetic nucleic acid obtained per single synthesis and reduces production efficiency

Method used

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  • Porous resin bead and production method of nucleic acid by using same
  • Porous resin bead and production method of nucleic acid by using same
  • Porous resin bead and production method of nucleic acid by using same

Examples

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preparation example Construction

[0064] The preparation method of the porous resin beads of the present invention is not particularly limited, and examples thereof include:

[0065] (1) A method comprising stirring a monovinyl monomer, a crosslinking vinyl monomer, a monovinyl monomer having a carboxyl group capable of reacting with a carboxyl group by a dehydration condensation reaction, in water (wherein the water contains a dispersion stabilizer dispersed or dissolved therein) Combined vinyl monomer, pore forming agent and polymerization initiator, and carry out suspension copolymerization,

[0066] (2) A method comprising stirring a monovinyl monomer, a crosslinking vinyl monomer, a vinyl monomer having other functional groups in water (wherein the water contains a dispersion stabilizer dispersed or dissolved therein) , a pore former and a polymerization initiator, suspension copolymerization is performed to obtain porous resin beads having other functional groups, and the other functional groups are conv...

Embodiment 1

[0088] (1) Preparation of porous resin beads

[0089] A 500 mL separable flask with a cooler, a stirrer, and a nitrogen inlet tube was placed on a constant temperature water bath, polyvinyl alcohol (manufactured by KURARAY CO., LTD., 2.6 g) and distilled water (260 g) were added, and the mixture was stirred at 300 rpm. mixture to dissolve polyvinyl alcohol. Added thereto by mixing styrene (manufactured by Wako Pure Chemical Industries, Ltd., 34.36 g, 71.6 mol% in the total monomer), p-acetoxystyrene (manufactured by Aldrich, 3.82 g, in the total monomer 5.1mol% in), divinylbenzene (a mixture of m-divinylbenzene and p-divinylbenzene, content 55wt%, manufactured by Wako Pure Chemical Industries, Ltd., 25.45g, 23.3 in the total monomer mol%), 2-ethylhexanol (manufactured by Wako Pure Chemical Industries, Ltd., 53.45 g), isooctane (manufactured by Wako Pure Chemical Industries, Ltd., 22.91 g) and benzoyl peroxide (containing 25 wt% water, manufactured by NOF CORPORATION, 1.27 g)...

Embodiment 2

[0104]In the same manner as in Example 1, except that styrene (20.16 g, 44.8 mol % in total monomers), p-acetoxystyrene (3.82 g, 5.5 mol % in total monomers) were used mol%), divinylbenzene (content 55wt%, 50.9g, 49.8mol% in the total monomer), 2-ethylhexanol (55.39g), isooctane (23.74g) and benzyl peroxide acyl (1.22 g) to obtain porous resin beads of a styrene-divinylbenzene-p-hydroxystyrene copolymer (the amount of hydroxyl groups per 1 g of porous resin beads calculated from the amount of monomers: 466 μmol / g).

[0105] Table 1 shows the maximum bead packing amount, median pore size, median particle size, and swelling volume in toluene of porous resin beads in a 0.2 mL column, which were measured in the same manner as in Example 1 and calculate.

[0106] In the same manner as in Example 1 and using the blending ratios shown in Table 2, porous resin beads bound to DMT-dT-3'-succinate were prepared, and DMT- dT-3'-succinate loading. The loadings are shown in Table 2.

[...

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Abstract

The present invention provides a porous resin bead of a copolymer composed of a monovinyl monomer unit and a crosslinking vinyl monomer unit, which has a group capable of binding with a carboxy group by a dehydration condensation reaction, wherein the amount of the crosslinking vinyl monomer is 18.5 - 55 mol% of the total monomer, and the median pore size of the porous resin bead is 60 - 300 nm.

Description

technical field [0001] The present invention relates to porous resin beads. The porous resin beads of the present invention can be used to prepare nucleic acids (especially polynucleotides of not less than 40 chain segments). Background technique [0002] A solid-phase synthesis method using the phosphoramidite method is widely used for the chemical synthesis of nucleic acids such as DNA oligonucleotides or polynucleotides and RNA oligonucleotides or polynucleotides. In this method, for example, a nucleoside which is the 3'-terminus of a nucleic acid to be synthesized is carried in advance by a solid-phase synthesis carrier via a cleavable linker (such as a succinyl group, etc.), and the carrier is placed on In the synthesis column, the synthesis column is placed on an automatic nucleic acid synthesizer. Thereafter, the synthesis reagents are flowed into the reaction column by the automatic nucleic acid synthesizer, and for example, the following synthesis procedure is car...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F212/08C08F212/14C08F212/36C08J9/28C07H21/04C07H1/00C40B50/18
CPCC08F8/12C07H21/00C07H1/00C08J2203/14C08J2325/08C08F212/22C08F212/36C08F212/08C08J9/141C08F12/22
Inventor 前田惠里森健二郎堀江将平
Owner NITTO DENKO CORP