Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Nucleic-acid extraction kit and application thereof

A kit and nucleic acid technology, applied in the field of molecular biology, can solve the problems such as the inability to extract a large amount of mortars, the inability to completely break the cell wall, and the inability to obtain a good wall removal effect, so as to be beneficial to removal, improve the lysis efficiency, and have a good effect. release effect

Inactive Publication Date: 2018-05-04
SUZHOU PRECISION BIOTECH CO LTD
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the mechanical method mainly uses liquid nitrogen freeze-drying to grind and break the cell wall, but this method cannot completely break the cell wall, and most of this method uses toxic organic solvents, phenol, etc., and the operation takes a long time, and it is easy to use liquid nitrogen. Frostbite skin, and limited by the number of mortars, it is impossible to extract a large amount
The enzymatic hydrolysis method to digest the cell wall also has the problem of not getting a good wall removal effect and low extraction efficiency

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Nucleic-acid extraction kit and application thereof
  • Nucleic-acid extraction kit and application thereof
  • Nucleic-acid extraction kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The preparation of embodiment 1 detection kit

[0055] (1) Preparation of reagents

[0056] Buffer A: 0.5-0.5M Tris-Cl buffer (pH5.0-8.0), 0.5-1.5M NaCl;

[0057] Enzyme solution B: 0.05-0.2M phosphate buffer (pH6.8-7.4), 0.25M MgCl2, 80-120mg / mL helicase;

[0058] Lysate C: 0.05-0.5M Tris-Cl buffer (pH5.0-8.0), 0.05-0.2M EDTA, 8-12% SDS, 8-12% Tween-40;

[0059] Suspension D: 5-40mg / ml RNAaseA

[0060] Remover E: 0.2-1M Al2(SO4)3, 80-350g / L CTAB, 0.5-1.5M NaCl;

[0061] Binding solution F: 0.1-0.5M Tris-Cl buffer solution (pH5.0-8.0), 2-8M guanidine isothiocyanate;

[0062] Wash solution G: 0.1-0.5M Tris-Cl buffer (pH5.0-8.0), 15-25% (volume ratio) absolute ethanol;

[0063] Eluent H: sterile deionized water;

[0064] (2) Consumables

[0065] Glass Beads, Spin Columns, and Collection Tubes

[0066] (3) Assembly of the kit

[0067] Assemble the primer composition, detection probe and reagents related to the kit to prepare the detection kit.

Embodiment 2

[0068] The extraction of embodiment 2 sample DNA (A1 / B1 / C1)

[0069] Select 3 stool samples: A, B, and C, divide them into 6 parts on average, and mark them as: A1-A6, B1-B6, C1-C6, and the three samples of A1, B1, and C1 are processed by the method of this embodiment. The DNA extraction method comprises the following steps:

[0070] (1) Take 800 μl of buffer A and 200 mg of glass beads into a 2 ml centrifuge tube;

[0071] (2) Add 200mg sample to the above 2ml centrifuge tube, vortex and mix for 30s;

[0072] (3) Add 100 μl enzyme solution B to the centrifuge tube in step (2), and incubate at 45-65°C for 20 minutes;

[0073] (4) Add 50 μl of lysate C to the sample, vortex for 5 minutes to mix the sample, centrifuge at 12,000 rpm (~13,400×g) for 60 s, transfer the supernatant (400 μl) to a new 2ml centrifuge tube;

[0074] (5) Add 300μl suspension D, vortex for 5-10s, place at room temperature for 5min, centrifuge at 12,000rpm (~13,400×g) for 60s, and precipitate the sample...

Embodiment 3

[0084] The extraction of embodiment 3 sample DNA (A1 / B1 / C1)

[0085] The three samples of A1, B1, and C1 are processed by the method of this embodiment, and the specific DNA extraction method includes the following steps:

[0086] (1) Take 800 μl of buffer A and 100 mg of glass beads into a 2 ml centrifuge tube;

[0087] (2) Add 200mg sample to the above 2ml centrifuge tube, vortex and mix for 30s;

[0088](3) Add 50 μl enzyme solution B to the centrifuge tube in step (2), and incubate at 40-50° C. for 120 minutes;

[0089] (4) Add 100 μl of Lysis Buffer C to the sample, vortex for 5 minutes to mix the sample, centrifuge at 12,000 rpm (~13,400×g) for 60 s, transfer the supernatant (400 μl) to a new 2ml centrifuge tube;

[0090] (5) Add 300μl suspension D, vortex for 5-10s, place at room temperature for 5min, centrifuge at 12,000rpm (~13,400×g) for 60s, and precipitate the sample particles;

[0091] (6) Transfer the supernatant to a new 2ml centrifuge tube, add 50μl remover ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the field of biology and relates to a nucleic-acid extraction kit and application thereof. The kit comprises glass beads, helicase, lysis solution and a remover, wherein the lysis solution comprises Tween-40, and the remover comprises CTAB and Al2(SO4)3. The kit has the advantages that fungal cell walls can be broken by high efficiency, and high-quality fungal nucleic acidcan be obtained in a fast and high-efficiency manner.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to a nucleic acid extraction kit and its application, in particular to a nucleic acid extraction kit for fungi and its application. Background technique [0002] Fungi are a large class of eukaryotic cell-type microorganisms. It is widely distributed in nature and has a wide variety. At present, there are 10,000 genera and more than 100,000 species. Most of them are beneficial to humans, while a few are harmful to humans and can cause diseases of humans, animals and plants. [0003] There are more than 400 species of fungi related to medicine, and 50-100 species are common, which can cause infectious, toxic and hypersensitivity diseases in humans. Such as: skin ringworm, tinea versicolor, pneumonia, enteritis, allergic dermatitis, etc. [0004] At present, the detection methods of common fungi in clinical practice are mainly direct microscopic examination and isolation culture metho...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/10C12N1/06C12Q1/6806C12Q1/04
CPCC12N1/06C12N15/1006C12Q1/6806C12Q2563/149
Inventor 穆延召朱永亮周燕
Owner SUZHOU PRECISION BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com