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Nucleic acid diluent

A diluent, nucleic acid technology, applied in the field of life analysis, can solve problems such as long-term storage, unstable standards or samples

Active Publication Date: 2021-10-29
SHANGHAI MYGT BIOPHARMACEUTICAL LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the standard or sample prepared in this way (especially the low concentration standard or sample) is often unstable and cannot be stored for a long time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: the stabilizing effect of diluent to standard substance

[0052] In this example, the stabilizing effect of the diluent of the present disclosure (taking 1×TE+F68+Lambda DNA as an example) on the standard (taking DNA standard as an example) was verified. Table 1 lists the composition of each dilution.

[0053] Table 1. Composition of Diluents

[0054]

[0055]

[0056] Use the dilutions numbered 1-7 above to perform the following serial dilutions on the DNA standard stock solution (10-fold serial dilution):

[0057] 1) 1v standard substance stock solution i+9v dilution solution to obtain standard substance solution ii;

[0058] 2) 1v standard solution ii+9v dilution to obtain standard solution iii;

[0059] 3) 1v standard solution iii+9v dilution to obtain standard solution iv;

[0060] 4) 1v standard solution iv+9v diluent to obtain standard solution v;

[0061] 5) 1v standard solution v+9v diluent to obtain standard solution vi;

[0062] 6)...

Embodiment 2

[0071] Embodiment 2: the stabilizing effect of diluent to nucleic acid sample

[0072] In this example, the stabilizing effect of the diluent of the present disclosure (taking 1×TE+F68+Lambda DNA as an example) on a nucleic acid sample (taking a DNA sample as an example) was verified. Table 2 lists sample recoveries using different dilutions.

[0073] Take an appropriate amount of the sample to be tested (the laboratory’s own virus sample S01201001), and add equal volumes of 120pg / μl, 60pg / μl and 30pg / μl standards respectively as the sample spike group. Both samples and standards were diluted with diluent 5 or a commercially available diluent (Shenke kit, diluent batch numbers 06301 and 0901).

[0074] Table 2. Sample recoveries

[0075]

[0076] The results show that the recovery rate of the dilution 5 (1×TE+F68+Lambda DNA) of the present disclosure is close to 100%, which is better and more reliable than the dilution of the commercially available kit.

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Abstract

The invention relates to a nucleic acid diluent. The nucleic acid diluent comprises 1*TE, F68 and DNA selected from the group consisting of Lambda DNA, pUC19 plasmid DNA, pUC18 plasmid DNA, pBR322 plasmid DNA, denatured protamine DNA, pUC57 plasmid DNA, pTZ19R DNA, Escherichia coli DNA and sssDNA. The nucleic acid diluent provided by the invention can be used for dilution of a standard substance and a sample in quantitative PCR and has the effects of protecting and stabilizing the standard substance and the sample. The invention also relates to a standard substance solution prepared from the nucleic acid diluent and a quantitative PCR method using the standard substance solution.

Description

technical field [0001] The present disclosure belongs to the field of life analysis. Specifically, the present disclosure relates to a nucleic acid diluent, which can be used for dilution of standards and samples in quantitative PCR. The present disclosure also relates to a standard solution prepared with the nucleic acid dilution and a quantitative PCR method using the standard solution. Background technique [0002] Quantitative PCR is a method that can measure the amount of PCR products, and can quantify the initial template copy number or concentration of a sample with high precision over a wide concentration range. Quantitative PCR has the advantages of simple operation, high sensitivity, and good repeatability. It has been widely used in various fields of life science research, such as differential expression analysis of genes, SNP detection, allele detection, drug development, clinical diagnosis, and transgenic research. Wide range of applications. [0003] Quantit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851
CPCC12Q1/6851C12Q2527/125C12Q2545/114
Inventor 王新涛蒋威郑静肖啸杜增民王利群赵阳吴侠陈晨郑浩
Owner SHANGHAI MYGT BIOPHARMACEUTICAL LLC
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