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Lysis solution for extracting nucleic acid through magnetic bead method

A technology for extracting nucleic acid and lysate, which is applied in the field of molecular biology, can solve the problems of low efficiency of lysate, and achieve the effect of authentic and reliable experimental results, good dispersion, and effective cell lysis

Active Publication Date: 2016-03-23
BEIJING NAGENE DIAGNOSTIC REAGENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] One aspect of the present invention is to provide a kind of lysate for magnetic bead method to extract nucleic acid for the shortcoming of low efficiency of lysate for nucleic acid extraction by magnetic bead method in the prior art

Method used

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  • Lysis solution for extracting nucleic acid through magnetic bead method
  • Lysis solution for extracting nucleic acid through magnetic bead method
  • Lysis solution for extracting nucleic acid through magnetic bead method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1: The real-time fluorescent quantitative PCR detection of HBVDNA

[0064] The reagents prepared according to the above-mentioned lysate scheme 1) and washing solution scheme 1) mixed with magnetic beads were used respectively for the clinical diagnosis and the tested HBVDNA quantitative value of 2×10 1 IU / ml hepatitis B patient's serum carries out the operation of following steps:

[0065] (1) Dispensing of PCR lysate: Dispense the prepared lysate mixed with magnetic beads into dedicated PCR tubes at a rate of 100 μl per tube.

[0066] (2) Adding samples: Take 100 μl of serum and add it to the above-mentioned PCR tube containing the lysate mixed with magnetic beads, gently blow and mix 5 times with a tip, and let stand at room temperature for 10 minutes.

[0067] (3) Aspirate and discard the liquid: place the above-mentioned PCR tubes on the eight-row magnetic stand, let it stand for 2 minutes, and use a pipette to absorb the liquid on the opposite side of ...

Embodiment 2

[0075] Embodiment 2: The real-time fluorescent quantitative PCR detection of HBVDNA

[0076] The reagents prepared according to the above-mentioned lysate scheme 2) and washing solution scheme 2) mixed with magnetic beads were used respectively for the clinical diagnosis and the tested HBVDNA quantitative value of 2×10 1 IU / ml hepatitis B patient's serum carries out the operation of following steps:

[0077] (1) Dispensing of PCR lysate: Dispense the prepared lysate mixed with magnetic beads into dedicated PCR tubes at a rate of 100 μl per tube.

[0078] (2) Adding samples: Take 100 μl of serum and add it to the above-mentioned PCR tube containing the lysate mixed with magnetic beads, gently blow and mix 5 times with a pipette tip, and let stand at room temperature for 5 minutes.

[0079] (3) Aspirate and discard the liquid: place the above-mentioned PCR tubes on the eight-row magnetic stand, let it stand for 2 minutes, and use a pipette to absorb the liquid on the opposite s...

Embodiment 3

[0087] Embodiment 3: The real-time fluorescent quantitative PCR detection of HBVDNA

[0088] Prepare the lysate and wash solution mixed with magnetic beads according to the following component ratios:

[0089] 0.3N Sodium Hydroxide, 0.45M Potassium Chloride, 0.03% Sodium N-Lauroyl Sarcosinate, 5mM EDTA, 0.45M Tris-HCL, 1% Triton X-100, 3mMDTT and 0.001wt% Bromophenol Blue.

[0090] Add the magnetic beads that have naturally precipitated for 24 hours according to the volume ratio of magnetic beads and lysate of 1:100, and mix well.

[0091] Washing liquid: 0.3M potassium chloride, 0.001wt% stone green, adjust the pH value to 4 with acetic acid.

[0092] Use the above lysate and wash solution to confirm the clinical diagnosis and the tested HBVDNA quantitative value is 2×10 1 IU / ml hepatitis B patient's serum carries out the operation of following steps:

[0093] (1) Dispensing of PCR lysate: Dispense the prepared lysate mixed with magnetic beads into dedicated PCR tubes at a...

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Abstract

The invention provides a lysis solution for extracting nucleic acid through a magnetic bead method. The lysis solution is prepared from 0.2-0.4 N sodium hydroxide, 0.3-0.6 M potassium chloride, 0.01-0.05% sodium n-lauroyl sarcosinate, 5mM EDTA, 0.3-0.6 M Tris-HCL and 1-2% triton X-100. The lysis solution for extracting nucleic acid through the magnetic bead method can fully and effectively lyse cells to accelerate the lysis progress and also can protect nucleic acid against oxidation to prevent formation of DNA dimmers. Besides, the lysis solution is stable in property and free of influences of seasons, temperature, saline ion concentration and the like and can carry out nucleic acid adsorption by fully cooperating with magnetic beads to achieve the optimal nucleic acid extraction effect. The scrubbing solution for extracting nucleic acid through the magnetic bead method can effectively remove residual impurities and avoid loss of nucleic acid, and a small quantity of the residual scrubbing solution does not influence the PCR application effect, so that the accuracy of detection results is guaranteed.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a lysate for extracting nucleic acid by a magnetic bead method. Background technique [0002] Nucleic acid is the carrier of genetic information, the most important biological information molecule, and the main object of molecular biology research. Therefore, nucleic acid extraction is the most important and basic operation in molecular biology experimental techniques. Nucleic acid extraction refers to the process of separating the nucleic acid from the carrier by physical and chemical methods. At present, most medical institutions and scientific research fields in China need to extract nucleic acid from a large number of blood samples. Therefore, in order to ensure that high-concentration nucleic acids are extracted, new technologies are constantly being developed. [0003] Nucleic acid extraction methods are various, including traditional phenol-chloroform method, salting-out...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 王海滨王棽周其玲冯小霞
Owner BEIJING NAGENE DIAGNOSTIC REAGENT CO LTD
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