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A nucleic acid extraction reagent and coronavirus technology, which is applied in the new coronavirus nucleic acid extraction kit and extraction field, can solve the problems of time-consuming, cumbersome and complicated operations, and achieve the effect of reducing costs and reducing the false negative rate
Active Publication Date: 2022-01-14
HANGZHOU ANYU TECH CO LTD
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Problems solved by technology
At present, the commonly used nucleic acid extraction technology has a complicated extraction process, a long time, and low nucleic acid extraction efficiency, which cannot meet the needs of clinical testing; commonly used viral nucleic acid extraction and purification technologies include Trizol method, spin column method, etc.
The Trizol method separates proteins and nucleic acids by using the different distribution positions of DNA, RNA, and proteins in the organic layer and the aqueous layer. The extraction of nucleic acids is cumbersome, and the reproducibility of the extraction effect for different samples is not good. It takes at least 1.5 h, the operation is cumbersome and time-consuming, so it is not suitable for clinical testing
The operation of conventional centrifugal adsorption column method for nucleic acid extraction is relatively complicated and time-consuming
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Embodiment 1
[0117] 1. Preparation of novel biological nanomagnetic beads
[0118] In parts by weight, 1 part of FeCl 3 ·6H 2 O, 0.38 parts of FeCl 2 4H 2 O and 5 parts of 1.5M hydrochloric acid solution were added with 1.12 parts of 1M ammonia water under stirring, magnetically separated after stirring and reacting for 35min, and the resulting solid matter was washed to obtain Fe 3 o 4 ;
[0119] By weight, 0.01 part of Fe 3 o 4 Mix with 0.5 parts of distilled water, then mix with 45 parts of isopropanol, add 1 part of PEG while shaking and stirring, continue stirring for 12 minutes, add 2.5 parts of distilled water and 1 part of TEOS, magnetically separate after stirring, and wash the obtained solid substance to obtain Fe 3 o 4 @SiO 2 nano magnetic beads;
[0120] React 1 part of urea with 1 part of chloromethyltrimethoxysilane under heat reflux for 3.5 hours, conduct vacuum distillation after the reaction, and collect the target fraction to obtain the modified silane coupling...
Embodiment 2
[0129] 1. Preparation of novel biological nanomagnetic beads
[0141] The composition and concentration of the novel coronavirus nucleic acid extraction kit are shown in Table 5 and Table 6.
[0142] Table 5 Components of the Novel Coronavirus Nucleic Acid Extraction Kit
[0143]
[0144] Table 6 The concentration of each component of the new coronavirus nucleic acid extraction kit
[0145]
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Abstract
The invention discloses a novel coronavirus nucleic acid extraction kit and a nucleic acid extraction method, belonging to the technical field of nucleic acid extraction. The kit comprises a suspension prepared from novel biological nano-magnetic beads. A preparation method of the novel biological nano-magnetic beads comprises the step of subjecting Fe3O4@SiO2 nano-magnetic beads to reacting with a modified silane coupling agent obtained by modifying chloromethyl trimethoxysilane with urea. The method for extracting novel coronavirus nucleic acid by using the kit comprises the following steps: adding a lysis solution into a sample, carrying out pyrolysis at room temperature, adding a washing solution, carrying out oscillating and centrifuging, taking supernate, adding the novel biological nano-magnetic bead suspension, and carrying out adsorbing at room temperature; after adsorption, removing supernate, adding a washing solution, and conducting uniform mixing; performing standing, removing supernate, and carrying out airing; and after airing, adding an eluent, and carrying out eluting at room temperature to obtain an RNA solution. The kit disclosed by the invention can be used for extracting high-purity and high-concentration nucleic acid, and the extracted nucleic acid is not easy to degrade.
Description
technical field [0001] The invention belongs to the technical field of nucleic acid extraction, in particular to a novel coronavirus nucleic acid extraction kit and extraction method. Background technique [0002] After the virus nucleic acid is extracted and purified and identified by PCR amplification, the test results can be used as one of the clinical references. However, due to the existence of exogenous and endogenous nucleases and their relatively stable enzyme activity, it is relatively difficult to extract and purify high-quality viral nucleic acids. The formula of nucleic acid extraction reagents has a great influence on the quality and yield of nucleic acid. Therefore, the development of efficient, rapid and high yield nucleic acid extraction reagents is a subject of continuous exploration in this field. As an automated nucleic acid extraction method, the magnetic bead method is based on the specific adsorption of magnetic beads on nucleic acid in high-salt solut...
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