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Method for storing nucleic acid specimens at room temperature, product and use thereof

A technology of room temperature preservation and nucleic acid, applied in the biological field, can solve the problems of space occupation, high price, unknown results, etc., and achieve the effect of simple preservation method, low cost and good yield

Active Publication Date: 2020-02-14
北京知因谱科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] First, low-temperature storage first requires a large number of refrigerators, which take up a lot of space. Secondly, both refrigerators and cold storage require a lot of power consumption, which is not only an economic burden, but also an energy burden, and a burden on environmental protection;
[0006] Second, there are relatively few reports on the use of liquid nitrogen to preserve nucleic acids, because liquid nitrogen is a consumable and needs to be replenished frequently, which makes this preservation method relatively expensive, and there are certain safety hazards;
[0007] Third, although DNA is very stable, it is also degrading itself over time. The theoretical upper limit of most reports is ten years
So if it is relatively pure nucleic acid, the result is unknown

Method used

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  • Method for storing nucleic acid specimens at room temperature, product and use thereof
  • Method for storing nucleic acid specimens at room temperature, product and use thereof
  • Method for storing nucleic acid specimens at room temperature, product and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preserving DNA at room temperature

[0032] This embodiment provides a method for storing nucleic acid samples at room temperature, including the following steps.

[0033] S1: Wash the silica microbeads, zirconia microbeads and carbon steel microbeads with acid, then mix 10 microbeads each, and put them in a 1.5mL centrifuge tube;

[0034] S2: Mix the extracted DNA with TE buffer to prepare DNA solutions respectively; add the DNA solutions to 1.5mL centrifuge tubes, place in a dry bath at 55°C for 24 hours, and mix 3 times in between;

[0035] S3: After being completely dried, store them separately at room temperature.

[0036] 4 kinds of microbeads with different DNA, respectively named TE-1 to TE-4 (the solvent is TE), the amount of DNA contained on each bead is about TE-1 (312ng); TE-2 (166ng); TE-3 (139ng); TE-4 (62-71ng).

[0037] After 3 days at room temperature, it was amplified with 8 pairs of different primers.

[0038] TE-1 to TE-3 each took out...

Embodiment 2

[0048] Example 2: Preserving Fresh Cells at Normal Temperature

[0049] This embodiment provides a method for storing nucleic acid samples at room temperature, including the following steps.

[0050] S1: Wash the silica microbeads, zirconia microbeads and carbon steel microbeads with acid, then mix 10 microbeads each, and put them in a 1.5mL centrifuge tube;

[0051] S2: fresh PB sample, according to the cell count, taken out containing 5 × 10 6 For the whole blood of white blood cells, use 5 times the volume of red blood cell lysate (0.5x) to lyse the red blood cells, and then resuspend them with 100 μL PBS; add the cell suspension to a 1.5 mL centrifuge tube, and dry in a vacuum oven;

[0052] S3: After completely drying, store in a dry place at room temperature, each bead contains about 1×10 5 white blood cells or an equivalent amount of DNA.

[0053] During the PCR reaction, 2 zirconia microbeads or 2 silica microbeads were added to each well, and 7 pairs of different p...

Embodiment 3

[0060] Example 3: Preserving cells treated with fixative at room temperature

[0061] (Cell) fixative is a necessary reagent for the preparation of karyotype analysis samples. It is prepared from methanol and glacial acetic acid at a ratio of 3:1. The fixed cell suspension (at this time, the most complete state of gDNA) can be Long-term storage at -30°C.

[0062] This embodiment provides a method for storing nucleic acid samples at room temperature, including the following steps.

[0063] S1: Wash the silica microbeads, zirconia microbeads and carbon steel microbeads with acid, then mix 10 microbeads each, and place them in 1.5mL centrifuge tubes;

[0064] S2: Take out 500 μl methanol-glacial acetic acid cell suspension (WBC total 2.5×10 6 ), placed in a 1.5ml EP tube, centrifuged at 5000rpm for 3min;

[0065] Suck out all the methanol-glacial acetic acid from the cell suspension, resuspend in PBS buffer (some loss of WBC), centrifuge at 8000rpm for 3min, suck out all the P...

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Abstract

The invention relates to a method for storing a nuclear acid sample at normal temperature, and a product and a using method thereof. The method for storing the nuclear acid sample at the normal temperature comprises the following steps: cleaning silicon dioxide micro-beads, zirconium oxide micro-beads and carbon steel micro-beads with an acid, and then mixing the same number of the silicon dioxide micro-beads, the zirconium oxide micro-beads and the carbon steel micro-beads; placing the mixed micro-beads in a container; adding the nuclear acid sample into the container; uniformly mixing, performing dry-bath or drying; and storing the obtained sample at room temperature. According to the method for storing the nuclear acid sample at the normal temperature provided by the invention, the nuclear acid is stored at the normal temperature for a long time by taking the surfaces of the micro-beads of a silicon dioxide material, a zirconium oxide material and the carbon steel material as carriers and utilizing the characteristics that the nuclear acid can be adsorbed with silicon dioxide substances in an incompatible manner and can be stored for a long time in a dry state; long-time storage of the nuclear acid can be realized by directly storing the DNA, and fresh and stationary liquid treated cells; the storage effect is good; and the storage method is simple.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for storing nucleic acid samples at room temperature, its product and its use method. Background technique [0002] With the rapid development of molecular biology detection technology, the detection of nucleic acid will play an important role in various fields. While testing, there are also a large number of nucleic acid samples that need to be preserved. These preserved nucleic acids may be used for scientific research, or play a greater role with the advancement of science and technology. [0003] Common nucleic acids include DNA and RNA. Among them, DNA is relatively stable and can be stored for a long time; RNA is extremely unstable. If it is to be stored for a long time, it needs to be reverse-transcribed into cDNA, and most laboratories only store it for 3 to 6 months. [0004] At present, cryopreservation methods are mostly used to store nucleic acid samples, such ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003C12N15/1013
Inventor 李坤
Owner 北京知因谱科技有限公司