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A kit for nucleic acid purification or fragment screening and its application method

A kit and nucleic acid technology, applied in biochemical equipment and methods, microbial measurement/inspection, DNA preparation, etc., can solve problems such as limited throughput of centrifuges, need for gel electrophoresis, inability to achieve high throughput, automation, etc.

Active Publication Date: 2018-08-07
BEIJING SYNSORBIO TECH CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are cumbersome and time-consuming, or require gel electrophoresis or centrifugation, and cannot achieve high-throughput and automated operations
The gel recovery method first needs to prepare the gel, electrophoresis, gel cutting, recovery and other processes, which consume a lot of time and cost a lot of manpower.
The spin column method generally requires steps such as adsorption, washing, and elution, during which a large number of centrifugation steps are required, each centrifugation is about 5 minutes, and the throughput of the centrifuge is limited, which limits the ease of operation. Of course, the above two methods for a small amount of samples It can be an option, but when batch operations need to be implemented, the above methods cannot satisfy

Method used

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  • A kit for nucleic acid purification or fragment screening and its application method
  • A kit for nucleic acid purification or fragment screening and its application method
  • A kit for nucleic acid purification or fragment screening and its application method

Examples

Experimental program
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Effect test

Embodiment 1

[0074] Nucleic acid purification or fragment screening kits

[0075] A nucleic acid purification or fragment screening kit, comprising a magnetic bead solution (i.e., an adsorption reagent), a washing reagent, and an eluting reagent; wherein,

[0076] Described magnetic bead solution comprises the silica carboxyl magnetic beads that concentration is 10ng / μL, the sodium chloride that concentration is 2.5mol / L, the PEG8000 that volume percentage concentration is 12%, the trihydroxymethane-hydrochloric acid (Tris-hydrochloric acid (Tris- HCL pH=6.0), store at 2°C-8°C.

[0077] The washing reagent contains ethanol with a concentration of 70% by volume, 100mmol / L sodium chloride, 10mmol / L ethylenediaminetetraacetic acid (EDTA), 10mmol / L trihydroxymethane-hydrochloric acid (Tris-HCLpH=8.0), now Ready to use, store at room temperature.

[0078] The elution reagent contained 10 mmol / L trihydroxymethane-hydrochloric acid (Tris-HCL pH=8.5), and was stored at room temperature.

[00...

Embodiment 2

[0107] Nucleic acid purification or fragment screening kits

[0108] A nucleic acid purification or fragment screening kit, comprising a magnetic bead solution (i.e., an adsorption reagent), a washing reagent, and an eluting reagent; wherein,

[0109] The magnetic bead solution comprises a concentration of 1ng / μL of silicon oxide carboxyl magnetic beads, a concentration of 0.1mol / L of sodium chloride, a volume percent concentration of 10% PEG8000, 10mol / L of trishydroxymethane-hydrochloric acid (Tris- HCL pH=6.0), store at 2°C-8°C.

[0110] The washing reagent contains 1% ethanol by volume concentration, 1mmol / L sodium chloride, 0.5mmol / L ethylenediaminetetraacetic acid (EDTA), 0.5mmol / L trihydroxymethane-hydrochloric acid (Tris-HCLpH=8.0) , Ready to use, store at room temperature.

[0111] The elution reagent contained 0.1 mmol / L trihydroxymethane-hydrochloric acid (Tris-HCL pH=8.5), and was stored at room temperature.

[0112] The nucleic acid purification method and th...

Embodiment 3

[0114] Nucleic acid purification or fragment screening kits

[0115] A nucleic acid purification or fragment screening kit, comprising a magnetic bead solution (i.e., an adsorption reagent), a washing reagent, and an eluting reagent; wherein,

[0116] Described magnetic bead solution comprises the silicon oxide carboxyl magnetic beads that concentration is 100ng / μL, the sodium chloride that concentration is 100mol / L, the volume percent concentration is the PEG8000 of 50%, the trihydroxymethane-hydrochloric acid (Tris-HCL) of 100mol / L pH=6.0), store at 2°C-8°C.

[0117] The washing reagent contains ethanol with a concentration of 85% by volume, 100mmol / L sodium chloride, 10mmol / L ethylenediaminetetraacetic acid (EDTA), 10mmol / L trihydroxymethane-hydrochloric acid (Tris-HCLpH=8.0), now Ready to use, store at room temperature.

[0118] The elution reagent contained 10 mmol / L trihydroxymethane-hydrochloric acid (Tris-HCL pH=8.5), and was stored at room temperature.

[0119] T...

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Abstract

The invention discloses a kit for nucleic acidpurification or fragment screening. The kit contains a magnetic bead solution, a washing reagent and an elution reagent, wherein the magnetic bead solution contains silica carboxyl magnetic beads with the concentration of 1-100ng / micro-L,sodium chloridewith the concentration of 0.1-100 mol / L, polyethylene glycol with the volume percentage of 1%-50% and a buffer solution. The invention also discloses a method for nucleic acidpurification or fragment screening by using the kit. The kit is used for nucleic acidpurification or fragment screening by using a magnetic bead method, is not limited by flux and can achieve high flux. In addition, automatic operation can be performed by cooperating with an automatic operating instrument based on the magnetic properties of magnetic beads.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit for nucleic acid purification or fragment screening and a use method thereof. Background technique [0002] High-throughput sequencing technology, that is, next generation sequencing technology (next generation sequencing) can sequence and quantify hundreds of thousands to millions of DNA molecules at a time, providing a powerful tool for basic biomedical research and clinical testing. And promote the scientific research and clinical transformation of genomics, which has established its cornerstone position in precision medicine. A lot of work needs to be done before a sample to be tested is sequenced by a high-throughput DNA sequencing machine, and the key step is library construction. Library construction includes processes such as nucleic acid fragmentation, repair reaction, A addition reaction, ligation reaction, fragment screening and purification, PCR amplifi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q1/6806C12Q2527/125C12Q2563/143C12Q2563/149
Inventor 崔晓锋
Owner BEIJING SYNSORBIO TECH CO LTD
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