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Actinomycete nucleic acid extraction detection reagent, kit and method and application thereof

A technology for detection kits and detection reagents, applied in biochemical equipment and methods, microbial measurement/inspection, DNA preparation, etc., can solve problems such as cross-contamination, long operation time, complicated steps, etc., and achieve cost reduction and time shortening , fast and efficient extraction effect

Active Publication Date: 2022-06-07
HUGOBIOTECH BEIJING CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the above two nucleic acid extraction techniques have complicated steps, especially the pretreatment steps involve multiple liquid phase transfers and solution addition processes, which makes the operation time of the whole process longer
Not only is it easy to cause cross-contamination, but also the cost is high, making the cost of large-scale and extensive application high.

Method used

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  • Actinomycete nucleic acid extraction detection reagent, kit and method and application thereof
  • Actinomycete nucleic acid extraction detection reagent, kit and method and application thereof
  • Actinomycete nucleic acid extraction detection reagent, kit and method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: a kind of actinomycetes nucleic acid extraction detection reagent

[0043] Actinomycetes nucleic acid extraction and detection reagent components are: InstaGene Matrix (1-5%, w / v, Bio-Rad), Tris-HCl (10-100mM, pH8.0), SDS (1-20%, w / v ), KCl (10-100 mM) and EDTA (1-10 mM) were mixed; the buffer was prepared, autoclaved and stored at 4°C for long-term use in separate devices.

Embodiment 2

[0044] Example 2: Actinomycete nucleic acid extraction and detection kit

[0045] Actinomycetes nucleic acid extraction and detection kit, the kit includes extraction and detection reagents, and the extraction and detection reagents contain InstaGene Matrix with a mass concentration w / v of 1%-5%; 10mmol / L-100mmol / L substance concentration It is Tris-HCl, pH is 8.0; mass concentration w / v is 1%-20% SDS; KCl substance concentration is 10mmol / L-100mmol / L; EDTA substance concentration is 1mmol / L-10mmol / L .

Embodiment 3

[0046] Embodiment 3: the method for extracting nucleic acid of Mycobacterium tuberculosis by nucleic acid extract

[0047] The nucleic acid extraction solution of the present invention carries out nucleic acid extraction of Mycobacterium tuberculosis, and the specific steps are as follows:

[0048] A: Centrifuge precipitation of 1ml of the prepared mixed bacterial solution, the parameter is 12000rpm, 1 minute, and remove the supernatant;

[0049] B: Add 200 μl of nucleic acid extraction solution to the colony precipitation, shake and mix; then place the mixture in a 100°C water bath for 8 minutes, and then cool at room temperature for 2 minutes;

[0050] C: high-speed centrifugation and precipitation of the lysate, the parameter is 12000rpm, 2 minutes;

[0051] D: Take 100 μl of supernatant for subsequent quality control and target gene detection;

[0052] Nucleic acid quality control. In this example, NanoReady (Hangzhou Suizhen) was used to control the quality of the extra...

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PUM

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Abstract

The invention provides an actinomycete nucleic acid extraction and detection reagent, a kit and a method and application thereof, the extraction and detection reagent contains InstaGeneMatrix, Tris-HCl, SDS, KCl and EDTA, the extraction and detection reagent can rapidly and efficiently extract mycobacterium tuberculosis nucleic acid DNA, and the extraction and detection limit of mycobacterium tuberculosis is increased to 8 copy number / ml; mycobacterium tuberculosis nucleic acid can be rapidly and efficiently extracted from a complex sample, and the potential clinical applicability of the extraction method is verified.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the technical field of extraction and detection of actinomycete nucleic acid. Background technique [0002] Tuberculosis (TB, tuberculosis) is a chronic bacterial disease caused by Mycobacterium tuberculosis complex (MTBC). found in other organizations. The disease can spread MTBC into the air through coughing, sneezing or spitting of individual patients, resulting in human-to-human transmission of tuberculosis, and the infection can invade multiple organs of the human body. According to statistics, about one-third of the global population is infected with MTBC, and there are as many as 10 million confirmed cases and 1.5 million deaths each year. In my country, TB is the second largest class A and B infectious disease after hepatitis B. To make matters worse, drug-resistant strains of MTBC are increasing, especially in developing countries. Therefore, how to accurately identify M...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12N15/1003C12Q1/689C12Q1/686C12Q1/6869C12Q2523/32C12Q2527/125C12Q2535/122C12Q2565/125Y02A50/30
Inventor 夏涵官远林付健尚芳娟王鹏军杨书恒李长诚佟斯垚
Owner HUGOBIOTECH BEIJING CO LTD
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