Solution for preserving and diluting DNA samples and preparation of solution

A diluent and sample technology, which is used in the preparation of DNA sample diluents, the dilution and preservation of ribonucleic acid samples in the field of polymerase chain reaction technology, and can solve the problem of short storage time at room temperature and inability to use the same buffer or Diluent, poor quality of the effect, etc., to achieve the effect of easy preservation

Pending Publication Date: 2016-04-27
GUANGZHOU BDS BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages are: 1) Different types of DNA samples (serum, plasma, tissue, urine, secretions, culture fluid, etc.) cannot use the same buffer or diluent; 2) Various buffers or diluents preserved The qu

Method used

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  • Solution for preserving and diluting DNA samples and preparation of solution
  • Solution for preserving and diluting DNA samples and preparation of solution
  • Solution for preserving and diluting DNA samples and preparation of solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: the preparation (100ml) of RNA dilution preservation solution

[0032] 1) Measure 30ml of deionized purified water and add it to a sterilized beaker;

[0033] 2) Add 1ml of dimethyl sulfoxide into the beaker, and stir evenly;

[0034] 3) Weigh 18.2g of sorbitol into the beaker, and stir to dissolve;

[0035] 4) Weigh 0.12g of Tris-HCl into the beaker, and stir to dissolve;

[0036] 5) Add 10ml ENDA (0.5mol / L) into the beaker and stir evenly;

[0037] 6) Add 0.5ml Triton-100 into the beaker and stir evenly;

[0038] 7) Add 10ml NP-40 (1%) into the beaker, and stir evenly;

[0039] 8) Weigh 1g of Chelex-100 into the beaker and stir evenly;

[0040] 9) Weigh 0.1g of sodium azide into the beaker, and stir to dissolve;

[0041] 10) Add 0.4ml of streptomycin (200U / ul) into the beaker, and stir evenly;

[0042] 11) Adjust the pH value to 7.8-8.2 with sodium hydroxide;

[0043] 12) Transfer the pH-adjusted solution to a 100ml volumetric flask, and dilute t...

Embodiment 2

[0045] Example 2: Secretion Sample Dilution Test

[0046] 1) Instruments, reagents and samples

[0047] Main instruments: ABI7500 PCR detector, TGL-18R high-speed refrigerated centrifuge, TDL-5-A desktop centrifuge, K10CD dry constant temperature metal bath, BSC-1500IIA2-X biological safety cabinet, pipette, etc.

[0048] Main reagents: Human papillomavirus (HPV) 16 and 18 nucleic acid amplification (PCR) fluorescent detection kits from Sun Yat-Sen University Daan Gene Co., Ltd.

[0049] Sample: fresh HPV16DNA clinically positive sample (secretion, 5.0E+07IU / ml).

[0050] 2) Experimental scheme

[0051] The HPV16 DNA positive samples were diluted 5 times, 50 times, 500 times and 5000 times with ordinary diluent and DNA sample dilution preservation solution respectively. Two sets of dilutions of the control group and the experimental group were designed, and the human papillomavirus (HPV) 16 and 18 nucleic acid amplification (PCR) fluorescence detection kits of Sun Yat-sen U...

Embodiment 3

[0060] Example 3: UU culture preservation test

[0061] 1) Instruments, reagents and samples

[0062] Main instruments: ABI7500 PCR detector, TGL-18R high-speed refrigerated centrifuge, TDL-5-A desktop centrifuge, K10CD dry constant temperature metal bath, BSC-1500IIA2-X biological safety cabinet, pipette, etc.

[0063] Main reagents: Ureaplasma urealyticum nucleic acid detection kit (PCR-fluorescent probe method) from Sun Yat-Sen University Daan Gene Co., Ltd.

[0064] Sample: UU culture (5.0E+07 IU / ml).

[0065] 2) Experimental scheme

[0066] Dilute the UUDNA culture 10 times and 1000 times with the DNA sample dilution preservation solution, and mark them as high and low concentration test samples as P1 and P2 respectively. Place them in -20±5°C environment, 2-8°C environment, room temperature (20-25°C) environment and 37°C environment respectively. The design of this experiment is as follows: mark the diluted samples and place them separately. In the designated environ...

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Abstract

The invention aims to provide a solution for preserving and diluting DNA samples, and belongs to the technical field of polymerase chain reaction (PCR) and the category of dilution and preservation of RNA (DNA) samples. The solution is prepared by the following steps: adding certain amounts of DNA preservative and chemical reagents into deionized purified water; adjusting the pH value to be 7.8-8.2, and enabling the volume to be constant; sealing and conducting autoclaved sterilization for later use at the room temperature. The solution is white and transparent, has the advantages of being stable, easy to preserve, convenient to use and the like, and is suitable for preserving and diluting the DNA samples of serum, plasma, tissue, urine, secreta and the like. When immersed into the solution for diluting and preserving the DNA samples, DNA in fresh tissue cells can be preserved for 1 week at 37 DEG C, 1 month at 25 DEG C, 6 months at 4 DEG C and long term at minus 20 DEG C to minus 80 DEG C in good condition.

Description

[0001] Field [0002] The invention belongs to the technical field of biological products, belongs to the dilution and storage category of ribonucleic acid (DNA) samples in the technical field of polymerase chain reaction (PCR), and relates to a preparation method and application of DNA sample diluents. Background of the invention [0003] Polymerase chain reaction (PCR) technology is an important molecular biology technique, which plays a very important role in the development of modern molecular biology. PCR technology is characterized by high sensitivity, high specificity, time-saving and rapidity. Samples for direct detection of target genes are divided into DNA samples or RNA samples, but the requirements for sample dilution and storage are very high. Ribonucleic acid (abbreviated as RNA, Ribonucleic Acid), the genetic information carrier present in biological cells and some viruses and viroids. RNA is a long chain molecule formed by condensation of ribonucleotides throu...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 李尔华高旭年钟凤然吴淑贤
Owner GUANGZHOU BDS BIOLOGICAL TECH CO LTD
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