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Porous plate with reagent-pre-storing micro-beads, preparation method and use method thereof

A technology of multi-well plates and reagents, which is applied in the field of high-throughput analysis and screening, and can solve the problems of cross-contamination of multi-well plates, easy deterioration of reagents, and large consumption of reagents, etc.

Active Publication Date: 2018-07-10
BEIJING DAWEI BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the existing pre-stored multi-well plates for reagents are stored in a dry state, the consumption of reagents is relatively large, the cost remains high, the sample volume required for screening reactions is large, the stored reagents are prone to deterioration, and the multi-well plates are easily separated. Cross-contamination occurs

Method used

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  • Porous plate with reagent-pre-storing micro-beads, preparation method and use method thereof
  • Porous plate with reagent-pre-storing micro-beads, preparation method and use method thereof
  • Porous plate with reagent-pre-storing micro-beads, preparation method and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Such as figure 1 , is a schematic diagram of the preparation method of the pre-stored reagent microbead multi-well plate. The multi-well plate used is a 96-well PCR plate, and a row of 96-well plates is shown in the figure, with a total of 8 containers; Ⅰ~Ⅷ refers to the diluted aqueous solution of screening reagents, and the dilution ratio is 40 times; the eight-channel pipette (1 ); pipetting reagent diluent (2); petroleum ether containing 0.01% Span80 (3); loaded reagent microbeads after water phase volatilization (4); mineral oil (5). The preparation method of the pre-storage reagent microbead porous plate, the specific steps are as follows:

[0027] a) Add 100 microliters of petroleum ether (containing 0.01% Span80) to each well of the 96-well plate in advance, and place the multi-well plate on a constant temperature metal bath at 4°C;

[0028] b) With an eight-way pipette (1-10 μl pipetting volume), draw 5 μL of the 40-fold dilution of the reagent to be screened...

Embodiment 2

[0035] Other conditions are the same as in Example 1, step e) is changed to add 100 microliters of mineral oil containing 6% petrolatum (liquid at room temperature). When this pre-stored reagent multi-well plate is stored at 4°C, the protective layer covering the beads will solidify. The position of the microbeads in the multi-well plate is fixed, and during storage and transportation, it can withstand unexpected vibration and flip without changing. When using a multi-well plate, the multi-well plate needs to be returned to room temperature, and the micro-sample injection operation is performed after the oil phase is melted.

Embodiment 3

[0037] Such as figure 2 Shown is a schematic flowchart of the microvolume screening method based on the multiwell plate of pre-stored reagent microbeads provided in Example 1 of the present application. The porous plate for pre-storing reagent microbeads comprises: a sealing film (1) for a sealed container, a porous plate (2), a first liquid (3), and reagent-loaded microbeads (4). The container of the porous plate used has a V-shaped bottom, so that microbeads or sample microdroplets converge at the center of the V-shape to achieve fusion, which is beneficial to the reaction screening. The first fluid used was mineral oil. The steps used include:

[0038] a) peel off the sealing film (1);

[0039] b) According to the micro-droplet generation method described in the published patent (Chinese patent, application number: 201410655191.5), add reagent or sample liquid to the first liquid (3) in the container of the porous plate through the micro-pipette device (5) drop(6);

...

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Abstract

The invention provides a reaction screening porous plate with reagent-pre-storing micro-beads, a preparation method and a use method thereof. According to the porous plate, the container of the porousplate is filled with a first liquid, reagent-loading micro-beads are pre-introduced into the bottom portion, the reagent-loading micro-beads can be solid microspheres, liquid micro-droplets, or any combination of solid microspheres and liquid micro-droplets, the first liquid is immiscible with the reagent-loading micro-beads, the porous plate is subjected to film sealing storage, and the reagentpreparation and the reagent adding and screening are not required by users during the use. According to the present invention, with the reagent-pre-storing micro-beads, the consumption of the reagentand the consumption of the sample can be substantially reduced, the mixing reaction with the subsequently-introduced trace sample can be conveniently performed, and the volume of the conventional screening reaction can be reduced by 2-3 orders of magnitude; and the reaction screening porous plate can be widely used for protein crystallization screening, drug interaction screening, microbial drug-resistance screening, culture condition screening, array PCR, single cell analysis and other fields.

Description

technical field [0001] The invention relates to the technical field of high-throughput analysis and screening, in particular to a micro-volume reaction screening porous plate based on pre-stored reagents. technical background [0002] Reaction screening and large-scale sample analysis and sorting in combinatorial chemistry, biology, clinical testing, and pharmacology mostly require manual preparation of reagents, which consumes a lot of reagents and consumables, and produces large errors during operation. Aiming at this bottleneck, the multi-well plate with pre-added reagents has been developed, which avoids the manual preparation of a large number of reagent solutions by users, is easy to use, and ensures the accuracy of reagent volume, which is of great significance. However, most of the existing pre-stored multi-well plates for reagents are stored in a dry state, the consumption of reagents is relatively large, the cost remains high, the sample volume required for screeni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48G01N35/00B01L3/02
CPCG01N33/48G01N35/00B01L3/02G01N2035/00465
Inventor 杜文斌奕巧莲
Owner BEIJING DAWEI BIOTECH LTD
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