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Preservation method for preparing trace amount of cell lysate in batches, and application thereof

A technology for micro-cell and storage methods, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve problems such as inter-well contamination, crystallization contamination, etc., to improve efficiency, improve lysate crystallization, and reduce cross-contamination. effect of chance

Active Publication Date: 2019-09-13
HANGZHOU BIOLYNX TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention aims at the problem that the cell or tissue lysate will produce crystallization under the condition of low temperature storage, which will cause contamination between pores, and provides a method for cell or tissue lysis and preservation. DMSO, or mixing glycerol and DMSO in different ratios and then adding it, solves the problem of crystallization and contamination

Method used

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  • Preservation method for preparing trace amount of cell lysate in batches, and application thereof
  • Preservation method for preparing trace amount of cell lysate in batches, and application thereof
  • Preservation method for preparing trace amount of cell lysate in batches, and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0049] (1) Add glycerin to Trizol or TCL buffer according to the ratio of 10%, 20%, 30%, 40%, 50% and 60% (v / v) and mix well;

[0050] (2) Add a certain volume of Trizol or TCL buffer lysate containing glycerol in the above ratio to a 96-well plate or a 384-well plate to lyse cells or tissues, and mix with a pipette or other tools if necessary;

[0051] (3) Quickly store the orifice plate of the cell or tissue lysate in a refrigerator at -20°C to -80°C;

[0052] (4) After storing for a period of time, take the cell or tissue lysate well plate out of the refrigerator, absorb the cell or tissue lysate after dissolving and carry out RT-PCR;

[0053] (5) Through electrophoresis detection, the bands are clear and free of miscellaneous bands, and the brightness is normal.

[0054] in Figure 2-5 Trizol or TCL buffer lysate after adding 10%, 20%, 30%, and 60% glycerol, respectively, no crystallization occurs after low temperature storage, indicating that Trizol or TCL buffer lysate a...

Embodiment 2

[0056] (1) Add DMSO to Trizol or TCL buffer according to the ratio of 10%, 20%, 30%, 40%, 50% and 60% (v / v) and mix well;

[0057] (2) Add a certain volume of Trizol or TCL buffer lysate containing glycerol in the above ratio to a 96-well plate or a 384-well plate to lyse cells or tissues, and mix with a pipette or other tools if necessary;

[0058] (3) Quickly store the orifice plate of the cell or tissue lysate in a refrigerator at -20°C to -80°C;

[0059] (4) After storing for a period of time, take the cell or tissue lysate well plate out of the refrigerator, absorb the cell or tissue lysate after dissolving and carry out RT-PCR;

[0060] (5) Through electrophoresis detection, the bands are clear and free of miscellaneous bands, and the brightness is normal. It shows that the Trizol or TCL buffer lysate after adding DMSO still maintains the original lysis effect and protection effect;

[0061] in Figure 6-9 Trizol or TCL buffer lysate after adding 10%, 20%, 30%, and 60...

Embodiment 3

[0063] (1) Add glycerin and DMSO at a ratio of 1:2 to 2:1, and then add them in proportions of 10%, 20%, 30%, 40%, 50% and 60% (v / v) in Trizol or TCL buffer and mix well;

[0064] (2) Add a certain volume of Trizol or TCL buffer lysate containing glycerol in the above ratio to a 96-well plate or a 384-well plate to lyse cells or tissues, and mix with a pipette or other tools if necessary;

[0065] (3) Quickly store the orifice plate of the cell or tissue lysate in a refrigerator at -20°C to -80°C;

[0066] (4) After storing for a period of time, take the cell or tissue lysate well plate out of the refrigerator, absorb the cell or tissue lysate after dissolving and carry out RT-PCR;

[0067] (5) Through electrophoresis detection, the bands are clear and free of miscellaneous bands, and the brightness is normal.

[0068] in Figure 10 Add 10% (DMSO:glycerol=1:2) TCL buffer cell lysate without crystallization after cryopreservation; Figure 11 Add 10% (DMSO: glycerol = 1: 1) ...

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Abstract

The present invention discloses a preservation method for preparing a trace amount of a cell lysate in batches. The preservation method comprises the following steps: (1) glycerol, DMSO or a mixture of the glycerol and DMSO are into a Trizol or TCL buffer lysate to be uniformly mixed; (2) the Trizol or TCL buffer lysate containing the additives is into a 96-well plate or a 384-well plate accordingto a certain volume to lyse cells or tissues, and a pipette is used for flowing and beating or other tools are used for uniform mixing; (3) the cell or tissue lysate well plates are rapidly placed ina refrigerator at a temperature of between -20 and 80 DEG C for preservation; (4) after stored for a period of time, the cell or tissue lysate well plates are taken out from the refrigerator, and thecell or tissue lysate is absorbed for RT-PCR after dissolution; and (5) electrophoresis detection is conducted. The method solves problems of crystallization and pollution by separately adding the glycerol in different proportions or the DMSO in different proportions or mixing the glycerol and DMSO in different proportions and then conducting adding.

Description

technical field [0001] The present invention belongs to the field of biotechnology; more specifically, the present invention reports a preservation method and its application for batch preparation of micro-cell lysates, so that nucleic acids can be subsequently extracted therefrom for the development of recombinant monoclonal antibodies. Background technique [0002] Nucleic acid is the core material in molecular biology research and the carrier of genetic information. Therefore, the preparation and preservation of nucleic acid is a key link in molecular biology experiments. Messenger RNA (mRNA) in nucleic acid is a type of single-stranded ribonucleic acid that carries genetic information and can guide protein synthesis. It plays an indispensable role in the transmission of genetic information, especially in the translation process from nucleic acid to protein. However, messenger RNA is very unstable due to its single-stranded form, and is easily polluted and degraded by RN...

Claims

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Application Information

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IPC IPC(8): C12N5/00
CPCC12N5/00
Inventor 李明振刘浩
Owner HANGZHOU BIOLYNX TECH CO LTD
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