Method for fluorescence determination in cell-free protein synthesis environment and porous plate

Abstract

The invention relates to a method for fluorescence determination in a cell-free protein synthesis environment and a porous plate. The method comprises the following steps: providing a porous plate, comprising a cover plate and a base provided with a plurality of holes, the holes being formed by one or more side walls, a bottom II and an opening, the cover plate being matched with the opening, andthe volume of a hole reaction cavity being smaller than 20 microliters; partial holes in the plurality of holes being communicated with one another; providing a fluid to part of the pores, and addingbiochemical factors and one or more of template DNA, template RNA, an additive and a reaction auxiliary factor when the fluid is a cell-free reaction mixture and a fluorescence measurement substance;when the fluid is a cell-free reaction mixture, a fluorescence measurement substance and a biochemical factor, adding one or more of template DNA, template RNA, an additive and a reaction auxiliary factor; placing a cover plate on the top of the base, and fluid making contact with the bottom II of the hole and the cover plate; incubating the porous plate for a period of time. The method and the porous plate can reduce reagent and determination cost.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for measuring fluorescence in a cell-free protein synthesis environment and a porous plate. Background technique [0002] Cell-free protein synthesis is also known as in vitro protein synthesis or CFPS. The aim of the process is to use the cell's biological machinery to produce proteins without being confined to living cells. Cell-free protein synthesis processes can sustainably produce proteins as long as there are sufficient concentrations of the reactive components. In general, cell-free protein synthesis requires the presence of amino acids, a DNA or RNA template encoding the desired protein, ribosomes, tRNA, and an energy source. Cell-free protein synthesis can be performed with purified single fractions or cell extracts. [0003] Fluorometric assays are often used in the context of cell-free protein synthesis, such as fluorescent protein detection. In such assays, ...

AI Technical Summary

Problems solved by technology

Although this plate can be widely used, there are following disadvantages in the field of this in vitro biological experiment: the volume provided by each well in the above-mentioned standard well plate is all large, such as in a standard 96-well plate, each well provides about 360 μL volume

For the above reaction using a 96-well plate with all wells, the cost of reagents can quickly rise to tens of thousands of yuan, and the cost of use is high.

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