Nucleic acid purification reagent based on magnetic bead method

A magnetic bead method and nucleic acid technology, applied in the field of molecular biology, can solve the problems of low recovery rate, difficulty in rare sampling, low initial amount of samples to be processed, etc., and achieve good recovery efficiency and low recovery loss

Inactive Publication Date: 2018-06-22
王煜
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If you encounter problems such as many steps in the operating system, difficult sampling of rare samples to be processed, and low initial volume of samples to be processed, how to efficiently perform nucleic acid purification and recovery in the experimental process pla

Method used

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  • Nucleic acid purification reagent based on magnetic bead method
  • Nucleic acid purification reagent based on magnetic bead method
  • Nucleic acid purification reagent based on magnetic bead method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Get the PCR purification products whose concentrations are respectively 16.6ng / μL (concentration one) and 22.8ng / μL (concentration two), contain the fragments that are 127bp and 306bp in size respectively, utilize nucleic acid purification reagent and kit of the present invention, measure two respectively The recoveries of samples with different concentrations.

[0019] 1) Put 20 μL of PCR purified product into a centrifuge tube, add 38 μL of nucleic acid purification reagent to the centrifuge tube, mix well, let stand at room temperature for 5 minutes, then place the centrifuge tube on a magnetic stand until the supernatant is clear, and discard the supernatant.

[0020] Wherein, the nucleic acid purification reagent is composed of the following raw materials in mass fractions: 5% carboxyl-modified magnetic beads, 19% sodium chloride, 18% polyethylene glycol, 0.04% sodium N-lauroyl sarcosinate, 0.16% Tris-HCl, 2% EDTA and 0.5% Tween 20, the balance is water.

[0021] ...

Embodiment 2

[0026] Get the PCR purification products whose concentrations are respectively 16.6ng / μL (concentration one) and 22.8ng / μL (concentration two), contain the fragments that are 127bp and 306bp in size respectively, utilize nucleic acid purification reagent and kit of the present invention, measure two respectively The recoveries of samples with different concentrations.

[0027] 1) Put 20 μL of PCR purified product into a centrifuge tube, add 38 μL of nucleic acid purification reagent to the centrifuge tube, mix well, let stand at room temperature for 5 minutes, then place the centrifuge tube on a magnetic stand until the supernatant is clear, and discard the supernatant.

[0028] Wherein, the components added in the nucleic acid purification reagent are: 3% carboxy-modified magnetic beads, 11.78% sodium chloride, 20% polyethylene glycol, 0.05% sodium N-lauroyl sarcosinate, 0.79% Tris-HCl , 0.5% EDTA, 1% Tween 20, and the rest is water.

[0029] 2) Add 200 μL of washing buffer ...

Embodiment 3

[0034] Get the PCR purification products whose concentrations are respectively 16.6ng / μL (concentration one) and 22.8ng / μL (concentration two), contain the fragments that are 127bp and 306bp in size respectively, utilize nucleic acid purification reagent and kit of the present invention, measure two respectively The recoveries of samples with different concentrations.

[0035] 1) Put 20 μL of PCR purified product into a centrifuge tube, add 38 μL of nucleic acid purification reagent to the centrifuge tube, mix well, let stand at room temperature for 5 minutes, then place the centrifuge tube on a magnetic stand until the supernatant is clear, and discard the supernatant.

[0036] Wherein, the components added in the nucleic acid purification reagent are: 1% carboxy-modified magnetic beads, 23.56% sodium chloride, 10% polyethylene glycol, 0.01% sodium N-lauroyl sarcosinate, 0.5% Tris-HCl , 1.5% EDTA, 2% Tween 20, and the remaining ingredients are water.

[0037]2) Add 200 μL of...

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Abstract

The invention relates to a nucleic acid purification reagent based on a magnetic bead method and a kit. The nucleic acid purification reagent is prepared from a water solution containing the followingsubstances in percentage by mass: 1 to 5 percent of carboxyl-modified magnetic beads, 11.78 to 23.56 percent of sodium chloride, 10 to 20 percent of polyethylene glycol, 0.01 to 0.05 percent of N-sodium lauroyl sarcosine, 0.16 to 0.79 percent of Tris-HCl, 0.5 to 2 percent of EDTA and 0.5 to 2 percent of tween-20. A nucleic acid purification recovery kit comprises the nucleic acid purification reagent, a washing buffer solution and an elution buffer solution. The nucleic acid purification reagent and the kit have the advantages that the nucleic acid recovery efficiency is higher than the recovery rate of the existing magnetic bead type reagent and kit; the recovery loss on trace samples is lower; the recovery efficiency on nucleic acid fragments being about 100bp or smaller nucleic acid fragments is better.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a nucleic acid purification reagent based on a magnetic bead method. Background technique [0002] At present, the purification of nucleic acid is inseparable from all fields of biomedicine. Many reactions, such as next-generation sequencing and TA cloning, require the purification and recovery of nucleic acid after enzymatic reactions. There are many ways to separate and purify nucleic acid: The concentrated salt method that dissolves different characteristics of RNA and RNA in a salt solution to separate the two, the organic solvent extraction method that uses organic solvents to denature proteins while inhibiting nuclease degradation, and the density of nucleic acids that are separated by using the principle of different densities of different contents Gradient centrifugation, as well as the adsorption material binding method that uses different adsorption materials (such as s...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 王龙罗喜鹏周甘霖李佩张青王煜
Owner 王煜
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