A kind of preservation method and application thereof for batch preparation of micro-cell lysate

A technology of trace cells and preservation methods, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve the problems of inter-well contamination, crystallization contamination, etc., to improve efficiency, improve lysate crystallization, and develop recombinant monoclonal antibodies. Process stability and mature effect

Active Publication Date: 2021-02-23
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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  • A kind of preservation method and application thereof for batch preparation of micro-cell lysate
  • A kind of preservation method and application thereof for batch preparation of micro-cell lysate
  • A kind of preservation method and application thereof for batch preparation of micro-cell lysate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] (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;

[0048](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;

[0049] (3) Quickly store the orifice plate of the cell or tissue lysate in a refrigerator at -20°C to -80°C;

[0050] (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;

[0051] (5) Through electrophoresis detection, the bands are clear and free of miscellaneous bands, and the brightness is normal.

[0052] 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 ...

Embodiment 2

[0054] (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;

[0055] (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;

[0056] (3) Quickly store the orifice plate of the cell or tissue lysate in a refrigerator at -20°C to -80°C;

[0057] (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;

[0058] (5) Through electrophoresis detection, the bands are clear and free of miscellaneous bands, and the brightness is normal. It shows that the Trizol or TCLbuffer lysate after adding DMSO still maintains the original lysis effect and protection effect;

[0059] in Figure 6-9 Trizol or TCL buffer lysate after adding 10%, 20%, 30%, and 60%...

Embodiment 3

[0061] (1) Add glycerin and DMSO in the ratio of 1:2 to 2:1, and then add them in the ratio of 10%, 20%, 30%, 40%, 50% and 60% (v / v) after mixing in Trizol or TCL buffer and mix well;

[0062] (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;

[0063] (3) Quickly store the orifice plate of the cell or tissue lysate in a refrigerator at -20°C to -80°C;

[0064] (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;

[0065] (5) Through electrophoresis detection, the bands are clear and free of miscellaneous bands, and the brightness is normal.

[0066] in Figure 10 Add 10% (DMSO:glycerol=1:2) TCL buffer cell lysate without crystallization after cryopreservation; Figure 11 Add 10% (DMSO: glyc...

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Abstract

The present invention is a kind of preservation method that is used for preparing trace cell lysates in batches, comprises the following steps: (1) adding glycerol, DMSO or the mixture of both into Trizol or TCL buffer lysates and mixing; (2) using Add a certain volume of Trizol or TCL buffer lysate containing the above additives to a 96-well plate or a 384-well plate to lyse the cells or tissues, and mix them evenly with a pipette or other tools if necessary; (3) Quickly dissolve the cells or tissues The orifice of the lysate should be stored in a refrigerator at -20°C to 80°C; (4) After a period of storage, take the cell or tissue lysate out of the refrigerator, absorb the cell or tissue lysate for RT-PCR after dissolving ; (5) detected by electrophoresis. The method solves the problems of crystallization and contamination by adding different proportions of glycerin or DMSO separately, or mixing glycerin and DMSO in different proportions and then adding them.

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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00
CPCC12N5/00
Inventor 李明振刘浩
Owner HANGZHOU BIOLYNX TECH CO LTD
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