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Cell preserving fluid and preparation method and use thereof

A technology of preservation solution and exfoliated cells, which is applied in the field of cell biology, can solve the problems that cell preservation solution cannot meet the needs of technological development, unsatisfactory DNA preservation, etc., and achieve the effect of preventing degradation

Active Publication Date: 2013-11-06
BGI GENOMICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the purpose of these preservation solutions is to use the preserved cells for cytological detection (such as TCT detection, that is, liquid-based thin-layer cytology detection), so they focus on fixing the cell structure and maintaining the shape of cells or proteins, but they have no effect on DNA. preservation is not ideal
With the development of technology, mass spectrometry detection and DNA sequencing have become important detection methods. The accuracy of detection results has higher and higher requirements for the collection, storage, and preparation of cell DNA samples. Cannot adapt to the needs of technological development

Method used

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  • Cell preserving fluid and preparation method and use thereof
  • Cell preserving fluid and preparation method and use thereof
  • Cell preserving fluid and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Preparation of Cell Preservation Solution 1

[0057] In Milli-Q ultrapure water, sequentially add the buffer solution and ionic strength maintainer in the formulation amounts shown in Table 1, adjust the pH to 8.0, and then add fixative, fixative auxiliary agent and rinse agent. Among them, the ratio of the phosphate buffer solution is: add sodium dihydrogen phosphate (NaH 2 PO 4 ·H 2 O) 0.9g, disodium hydrogen phosphate (Na 2 HPO 4 ·7H 2 O) 24.97g.

[0058] Table 1: Recipe of Cell Preservation Solution 1

[0059] component name

[0060] Those skilled in the art can understand that although the weight unit of each component in the above Table 1 is g or ml, it can also be understood as parts by weight or parts by volume respectively, that is, each component meets the requirements in the above Table 1. The ratio will do.

Embodiment 2

[0061] Embodiment 2: Preparation of Cell Preservation Solution 2

[0062] Cell preservation solution 2 was prepared according to the same method as in Example 1, except that the components used and their addition amounts were shown in Table 2 below. The ratio of the acetic acid buffer solution is: add acetic acid (CH 3 COOH) 0.32g, sodium acetate (CH 3 COONa) 12.89 g).

[0063] Table 2: Recipe of Cell Preservation Solution 2

[0064] component name

[0065] Those skilled in the art can understand that although the weight unit of each component in the above Table 2 is g or ml, it can also be understood as parts by weight or parts by volume respectively, that is, each component meets the requirements in the above Table 2 The ratio will do.

Embodiment 3

[0066] Embodiment 3: Preparation of Cell Preservation Solution 3

[0067] Cell preservation solution 3 was prepared according to the same method as in Example 1, except that the components used and their addition amounts were shown in Table 3 below. Wherein the piperazine-N, N'-bis(2-ethanesulfonic acid) (PIPES) buffer solution has a ratio of: 3.02 g of piperazine-N, N'-bis(2-ethanesulfonic acid) is added to 150 ml of water.

[0068] Table 3: Components and amounts of cell preservation solution 3

[0069] component name

[0070] Those skilled in the art can understand that although the weight unit of each component in the above Table 3 is g or ml, it can also be understood as parts by weight or parts by volume, respectively, that is, between the components that meet the requirements in the above Table 3 The ratio will do.

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Abstract

The invention belongs to the field of cell biology, relating to a cell preserving fluid and a preparation method and use thereof. Concretely, the cell preserving fluid comprises a fixing agent, a fixing agent assistant, an anticoagulant, a cushion fluid and an ionic strength maintenance agent. The cell preserving fluid is characterized in that the content of the anticoagulant is 0.01% to 1.5%(w / w); the content of the ionic strength maintenance agent is 0.01% to 1%(w / w); and the cell preserving fluid does not contain a disulfide bond open reagent. The cell preserving fluid can effectively preserve cell DNA (deoxyribonucleic acid) and virus DNA infecting the cell, is convenient to collect the cast-off cells and extract DNA and can give attention to storage of the cell structure. The invention also relates to the preparation method and the use of the cell preserving fluid. The invention also relates to a kit and a method for detecting HPV (Human Papilloma Virus) or HPV DNA.

Description

technical field [0001] The invention belongs to the field of cell biology, and relates to a cell preservation solution, a preparation method and application of the cell preservation solution. The present invention also relates to kits and methods for detecting HPV or HPV DNA. Background technique [0002] At present, most of the cell preservation solutions sold on the market are cell preservation solutions with alcohols, anticoagulants, buffer solutions, etc. as the main components. For example: [0003] European Patent Application Publication No. EP0511430A2 discloses a cell preservation solution, the main components of which are alcohols, acetic acid, and salt ions. Among them: alcohols (45%-55%), mainly methanol, ethanol and isopropanol; anticoagulants (2-4%), mainly EDTA and its salts; buffer solutions (6%-8%), for Phosphate buffer, sodium acetate buffer, citrate buffer, etc. [0004] The Chinese patent application with publication number CN101363011A discloses a cer...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/07
Inventor 易吉
Owner BGI GENOMICS CO LTD
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