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RNA extraction kit suitable for normal-temperature transportation, and extraction method

A kit and normal temperature technology, applied in the field of nucleic acid processing, can solve problems such as inability to transport at normal temperature, and achieve the effect of accelerating digestion speed, enhancing removal ability, and simple operation process

Active Publication Date: 2021-04-16
苏州赛普生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above-mentioned shortcomings and deficiencies of the prior art, the present invention provides an RNA extraction kit suitable for transportation at room temperature, which solves the problem that proteinase K-based RNA extraction kits cannot be transported at room temperature, and the RNA extraction kit of the present invention The kit does not contain toxic organic solvents and does not require heat treatment during the extraction of sample RNA. It uses one-step processing to complete lysis and binding at the same time. It is not necessary to separate lysis and binding. It is more suitable for automatic nucleic acid extraction instruments. The operation process is simple and fast, and it is also applicable Since the reagents are pre-packed into deep-well plates, the automated operation is easier and faster

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  • RNA extraction kit suitable for normal-temperature transportation, and extraction method
  • RNA extraction kit suitable for normal-temperature transportation, and extraction method
  • RNA extraction kit suitable for normal-temperature transportation, and extraction method

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Embodiment

[0049] Adopt the RNA virus nucleic acid extraction kit (Cellpro) of the present invention and the RNA virus nucleic acid extraction kit (CompA) on the market to carry out RNA pseudovirus nucleic acid extraction, and detect through reverse transcription, fluorescent quantitative PCR detection kit.

[0050] Sample preparation: RNA pseudovirus concentration is 5000copies / mL, 500copies / mL sample, 8 copies of each concentration of each nucleic acid extraction kit are extracted in parallel, and nucleic acid extraction is performed according to the extraction procedures in the respective kit instructions. After extraction, reverse transcription and qPCR detection were performed.

[0051] Wherein, the composition of RNA virus nucleic acid extraction kit (Cellpro) of the present invention is as follows:

[0052] ① Lysis binding solution, the composition is as follows (Table 1):

[0053]

[0054]

[0055] ②Magnetic bead suspension, the composition is as follows:

[0056] Superpa...

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Abstract

The invention relates to an RNA extraction kit suitable for normal-temperature transportation. The kit comprises a lysis binding solution, and the lysis binding solution contains guanidine isothiocyanate with the concentration of 1 to 5 M, sodium thiocyanate with the concentration of 0.2 to 3 M, trihydroxymethyl aminomethane with the concentration of 0.01 to 0.1 M, tetrasodium iminodisuccinate with the concentration of 0.001 to 0.05 M, sodium ascorbate with the mass fraction of 0.05-1%, betaine with the mass fraction of 0.1-2%, Tween 20 with the mass fraction of 1-6%, sodium acetate with the concentration of 0.05 to 0.5 M, and isopropanol withthe mass fraction of 25-55%. The lysis binding solution does not contain toxic organic solvents such as phenol and does not contain reagents such as protease K needing cold-chain transportation, so that the RNA extraction kit can be transported under the condition of normal temperature. The lysis binding solution has the effects of lysis of cells / viruses and promotion of combination of nucleic acid and magnetic beads, can complete lysis and combination in one step, and is more suitable for an automatic nucleic acid extractor. The two protein denaturants are combined for use, and compared with single use of one protein denaturant, the capacity of removing impurities such as protein with complex chemical properties can be enhanced.

Description

technical field [0001] The invention relates to the technical field of nucleic acid processing, in particular to an RNA extraction kit and extraction method suitable for normal temperature transportation. Background technique [0002] With the development of detection technology, pathogenic nucleic acid detection has become the mainstream detection method for the diagnosis of respiratory pathogens due to its high sensitivity and high specificity detection performance. By detecting the nucleic acid of pathogens in respiratory specimens such as nasopharyngeal swabs, airway extracts or sputum, it can accurately and quickly detect and distinguish common respiratory system infections from malignant pathogenic infections, and make rapid diagnosis of pathogenic infections, and can quickly Distinguish the pathogen type and subtype / strain, and provide basis and guidance for subsequent targeted prevention and treatment measures. [0003] Nucleic acid extraction is the first step in p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 张胜有叶竹青
Owner 苏州赛普生物科技股份有限公司
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