Urine sample RNA stabilizing solution and preparation method thereof

A urine sample and stabilizing solution technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as the inability to preserve urine RNA for a long time, achieve convenient later production and application, slow down decomposition, and improve efficiency and detection effect

Pending Publication Date: 2020-02-07
HANGZHOU YORK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the problem that the existing urine sample RNA stabilizing solution cannot preserve urine RNA for a long time, provide a urine sample RNA stabilizing solution, which can effectively maintain the stability of RNA in the urine sample, and realize large-volume urine Liquid storage and transportation, and suitable for subsequent RNA extraction and fluorescent RT-PCR

Method used

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  • Urine sample RNA stabilizing solution and preparation method thereof
  • Urine sample RNA stabilizing solution and preparation method thereof
  • Urine sample RNA stabilizing solution and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0028] The above description is only an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention, it can be implemented according to the contents of the description, and in order to make the above and other purposes, features and advantages of the present invention more obvious and understandable , the specific embodiments of the present invention are enumerated below.

[0029] The present invention provides a urine sample RNA stable solution, which is used to solve the problem that the existing urine sample RNA stable solution cannot preserve urine RNA for a long time and cannot realize the preservation and transportation of large volumes of urine. In order to solve the above problems, the present invention The general train of thought of invention is as follows:

[0030] Step 1: Configure AmL ultrapure water;

[0031] Step 2: Weigh dithiothreitol, guanidine isothiocyanate, and ethylenediaminetetr...

Embodiment 2

[0044] Prepare the urine sample RNA stabilization solution, and control the content of each component: dithiothreitol 0.01M, guanidine isothiocyanate 2M, diethyl pyrocarbonate 0.05mM, ethylenediaminetetraacetic acid 200mM; according to the above steps to configure stable liquid. In the same urine sample, 90 mL of urine was taken respectively. The experimental group was added with 10 mL of RNA stabilization solution, and the control group was added with 10 mL of ultrapure water. , 5 and 7 days later, use the TRIZOL method to extract urine total RNA, and use NanoDrop to measure the RNA content. The results are shown in Table 2:

[0045]

[0046] Table 2

Embodiment 3

[0048] The urine sample RNA stabilization solution was prepared, and the content of each component was controlled as follows: dithiothreitol 2M, guanidine isothiocyanate 6M, diethyl pyrocarbonate 0.5mM, and ethylenediaminetetraacetic acid 100mM. Follow the steps above to prepare the stabilizing solution. In the same urine sample, 90 mL of urine was taken respectively. The experimental group was added with 10 mL of RNA stabilization solution, and the control group was added with 10 mL of ultrapure water. , 5 and 7 days later, use the TRIZOL method to extract urine total RNA, and use NanoDrop to measure the RNA content. The results are shown in Table 3:

[0049]

[0050] table 3

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PUM

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Abstract

The invention discloses a urine sample RNA stabilizing solution and a preparation method thereof. The urine sample RNA stabilizing solution is prepared from 0.01 to 2M of dithiothreitol, 2M to 6M of guanidinium isothiocyanate, 1 to 200 mM of ethylenediamine tetraacetic acid, 0.05 to 0.5 mM of diethyl pyrocarbonate and ultrapure water. The preparation method comprises the following steps: adding acertain amount of the chemical reagent into ultrapure water, adjusting the pH value to 4-4.2 with hydrochloric acid, fixing the volume, and carrying out sealed storage. The urine sample RNA stabilizing solution provided by the invention can effectively maintain the stability of RNA in a urine sample, can realize preservation and transportation of large-volume urine, and is suitable for subsequentRNA extraction and fluorescent RT-PCR.

Description

technical field [0001] The invention relates to an RNA sample stabilizing liquid, in particular to a urine sample RNA stabilizing liquid and a preparation method thereof. Background technique [0002] Liquid biopsy is a technology that uses human body fluids (blood, urine and other body fluids) as a source of specimens to detect and obtain tumor-related information. It is a simple and non-invasive alternative to tissue biopsy. It has the advantages of good patient compliance, good specificity, low heterogeneity, and repeated sampling. Therefore, the early screening of cancer by liquid biopsy technology has very good development prospects. [0003] Urine is produced by the kidneys, stored in the bladder via the ureters, and then excreted through the urethra. Due to the physiological characteristics of urine, urine samples are an excellent source of liquid biopsy samples for urinary system tumors. Therefore, urinary system tumors, such as bladder cancer and prostate cancer, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2527/125C12Q2527/127C12Q2523/113
Inventor 朱友杰孔繁平
Owner HANGZHOU YORK BIOTECH CO LTD
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