Reagent composition for stabilizing nucleic acid molecules and application thereof

A nucleic acid molecule and composition technology, applied in recombinant DNA technology, DNA preparation, etc., can solve problems such as damage results

Pending Publication Date: 2021-05-07
苏州白垩纪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has been found that nucleic acids in biological samples may compromise the results of subsequent analyzes if not properly preserved and stabilized. If prec

Method used

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  • Reagent composition for stabilizing nucleic acid molecules and application thereof
  • Reagent composition for stabilizing nucleic acid molecules and application thereof
  • Reagent composition for stabilizing nucleic acid molecules and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Embodiment 1: Saliva sample DNA preservation reagent composition

[0091] The reagent composition is a combination of a pH stabilizer, a chelating agent, a protein denaturing agent, an antimicrobial agent, an inorganic salt ion and a reducing agent. Adopt deionized water to prepare DNA preservation reagent composition (S1) according to the following components and concentrations: 100mM sodium acetate, 100mM sodium citrate, 50mM alanine, 25mM sodium tartrate, 2.5M urea, 0.1% octane polyoxyethylene Dodecyl ammonium chloride, 10% ethanol, 10mM sodium sulfite, pH 9.5.

[0092] Adopt deionized water to prepare DNA preservation reagent composition (S2) according to following composition and concentration: 100mM sodium acetate, 100mM sodium citrate, 50mM alanine, 50mM EDTA, 2% sodium lauryl sulfate, 0.5% polyethylene glycol Glycol octyl phenyl ether, 10 mM sodium sulfite, 0.05% gentamicin, pH 9.0.

[0093] The subject rinsed his mouth with cold water three times, then waited...

Embodiment 2

[0111] Embodiment 2: swab sample DNA preservation reagent composition

[0112] The reagent composition is a combination of a pH stabilizer, a chelating agent, a protein denaturing agent and an antimicrobial agent.

[0113] Prepare the reagent composition with deionized water according to the following components and concentrations: 100mM sodium acetate, 100mM sodium citrate, 50mM alanine, 50mM trioxalic acid, 0.5% sodium lauryl sulfate, 0.2% NP40 and 0.05% Qingda Mycin. The pHs were 5, 6, 7, 8, 9, 10, 11 and 12, respectively.

[0114] The subjects rinsed their mouths with cold water three times, and then waited for 5 minutes to scrape oral swabs with flocked swabs according to the standard. After taking 8 swabs, add 1 mL of each of the above preservation solutions, mix well at room temperature, and place them for 1 month. The above-mentioned prepared samples were extracted using CretBiotech's kit "Magnetic Bead Method Saliva & Swab Genomic DNA Extraction Kit" (Cat#CNA004901)...

Embodiment 3

[0118] Embodiment 3: plasma sample viral DNA preservation reagent composition

[0119] The reagent composition is a combination of a pH stabilizer, a chelating agent, a protein denaturing agent, an antimicrobial agent, an inorganic salt ion and a reducing agent.

[0120] Adopt deionized water to prepare microbial DNA preservation reagent composition according to following composition and concentration: 100mM sodium acetate, 100mM sodium citrate, 50mM alanine 50mM trioxalic acid, 0.5% sodium lauryl sulfate, 0.1% octane poly Oxyethylene lauryl ammonium chloride, 0.05% gentamicin, 100 mM lithium chloride, 10 mM sodium sulfite, pH 10, and this formulation was labeled SB10.

[0121] Dilute the virus-containing positive plasma sample with negative plasma, or use the market's conventional VTM (hanks solution-based virus preservation solution) or the above-mentioned reagent combination SB10 for dilution, the dilution ratio is 1:1, and after mixing, place it for a few days. The viral ...

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Abstract

The invention provides a reagent composition for stabilizing nucleic acid molecules in a biological sample. The reagent composition comprises a pH stabilizer, a chelating agent, a protein denaturant, an antimicrobial agent and optional inorganic salt ions, a tissue penetrant, a reducer, a nuclease inhibitor and the like. The invention also provides a method for transporting, storing and stabilizing the nucleic acid molecules in the biological sample by using the reagent composition and application of the reagent composition in transporting, storing and stabilizing the nucleic acid molecules in the biological sample. The reagent composition ensures that high molecular weight DNA in the biological sample is kept for a long time at normal temperature.

Description

technical field [0001] The present invention relates to the technical field for stabilizing nucleic acid molecules in biological samples. More specifically, the present invention relates to reagent compositions and uses thereof for stabilizing nucleic acid molecules present in biological samples. Background technique [0002] In the field of diagnostics, nucleic acids are important biomarkers. For example, genes comprising the human genome or fragments thereof are widely used as biomarkers in molecular in vitro diagnostics to provide insights into normal biological and pathological processes and to predict disease outcomes and guide precision medicine. Therefore, nucleic acid molecules are important in disease diagnosis, prognosis, and clinical trials for biomarker discovery. Genomics is a powerful tool for exploring basic biology, diagnosing diseases, facilitating drug development, tailoring therapies for specific pathological and genetic conditions, and generating databa...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/10
Inventor 张佳斌何宗顺邹永龙曲峰
Owner 苏州白垩纪生物科技有限公司
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