Preparation method of micron-sized biological magnetic beads for purifying free DNA

A micron-scale biomagnetic technology, applied in the combination of nanomaterials and biotechnology engineering, can solve the problems of uneven particle size, small specific surface area, poor static suspension, etc., and achieve large nucleic acid binding and low salt residue , Combined with the effect of high efficiency

Inactive Publication Date: 2020-08-21
上海百英生物科技有限公司
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0004] However, there are currently the following problems: the price of imported magnetic beads is high, most of the domestic magnetic beads have a submicron particle size, the particle size is not uniform, the specific surface area is small, the static suspension is poor, the wall is sticky, and the redispersibility is poor.
As a result, the ability to bind to small fragments of free DNA in plasma is weak, and it is easy to lose during the re-extraction process, resulting in low yield and reduced detection sensitivity

Method used

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  • Preparation method of micron-sized biological magnetic beads for purifying free DNA
  • Preparation method of micron-sized biological magnetic beads for purifying free DNA
  • Preparation method of micron-sized biological magnetic beads for purifying free DNA

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preparation example Construction

[0065] The embodiment of the present invention provides a method for preparing micron-scale biological magnetic beads for purifying free DNA. It is characterized in that it includes the following steps:

[0066] Step 1: Synthesize polystyrene microspheres or polymethyl methacrylate microspheres as particle size template microspheres by dispersion polymerization;

[0067] Measure 100-2000mL absolute ethanol, 2%-10% (v / v, ultrapure water / (absolute ethanol + ultrapure water))d ultrapure water, 2%-8% (g / 100ml, Dispersant / (anhydrous ethanol + ultrapure water)) polyvinylpyrrolidone (average molecular weight 58K) into a 250-2000mL three-necked flask, stir until the solution is clear, pass high-purity nitrogen for 10-30min to remove oxygen in the solution, add 5%- 12% (g / 100ml, polymerized monomer / (anhydrous ethanol + ultrapure water)) styrene / methyl methacrylate which removes water and polymerization inhibitor after vacuum distillation, 0.5%-3%(g / g , Crosslinking agent / polymerization mo...

Embodiment 1

[0075] A method for preparing micron-scale biological magnetic beads for purification of free DNA. It is characterized in that it includes the following steps:

[0076] Step 1: Synthesize polystyrene microspheres as particle size template microspheres by dispersion polymerization;

[0077] Measure 1400mL absolute ethanol, 100mL ultrapure water, 50g polyvinylpyrrolidone (average molecular weight 58K) into a 2L three-necked flask, stir until the solution is clear, pass high-purity nitrogen for 15min to remove oxygen in the solution, add 120g vacuum distillation and remove Water removes the styrene of the polymerization inhibitor, 2.4g divinylbenzene, 3.6g initiator azobisisobutyl cyanide, the speed is set to 150rpm, stirred for a few minutes until the mixture is clear, the water bath is heated to 70°C, and react 24 hour. After the reaction, it was cooled to room temperature and washed with ethanol and water by centrifugation for 3 times. The centrifugal speed was 9000 rpm and the c...

Embodiment 2

[0086] A method for preparing micron-scale biological magnetic beads for purification of free DNA. It is characterized in that it includes the following steps:

[0087] Step 1: Synthesize polystyrene microspheres as particle size template microspheres by dispersion polymerization;

[0088] Measure 1400mL absolute ethanol, 80mL ultrapure water, 35g polyvinylpyrrolidone (average molecular weight 58K) into a 2L three-necked flask respectively, stir until the solution is clear, pass in high-purity nitrogen for 15min to remove oxygen in the solution, add 120g of reduced pressure distillation and remove Water removes the styrene of the polymerization inhibitor, 2.4g divinylbenzene, 3.6g initiator azobisisobutyl cyanide, the speed is set to 150rpm, stirred for a few minutes until the mixture is clear, the water bath is heated to 70°C, and react 24 hour. After the reaction, it was cooled to room temperature, washed with ethanol and water by centrifugation for 3 times, the centrifugal spe...

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Abstract

The invention discloses a preparation method of micron-sized biological magnetic beads for purifying free DNA. The method is simple in process, and the prepared magnetic beads are of a core-shell structure, uniform in particle size, monodisperse, free of agglomeration, high in magnetic response performance, rough in surface, rich in morphology, large in specific surface area and rich in surface hydroxyl content. The magnetic beads can be used for purifying free DNA samples.

Description

Technical field [0001] The invention belongs to the combined field of nanomaterials and biotechnology engineering, and specifically relates to a preparation method of micron-level biological magnetic beads for purifying free DNA. Background technique [0002] Traditional methods for separating nucleic acid mainly include salting-out method, silica method, organic method, etc., which are time-consuming and labor-intensive and use organic solvents. [0003] Biomagnetic beads are a new type of functional material involving nanomaterial science and biological science. The surface is easy to modify, solid-liquid separation can be quickly realized by applying a magnetic field, and the operation is simple. Biomagnetic beads have good application prospects in the extraction of trace nucleic acids and cell screening. Fast and efficient isolation of high-purity nucleic acids is an important prerequisite for various scientific researches in the field of modern molecular biology. [0004] How...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10B01J13/14
CPCC12N15/1013B01J13/14
Inventor 陆圣珏谭建雄郑飞剑查长春程千文
Owner 上海百英生物科技有限公司
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