Multiplex fluorescent PCR universal freeze-drying protective agent and application thereof

Abstract

The invention discloses a multiplex fluorescent PCR universal freeze-drying protective agent and an application thereof. The freeze-drying protective agent consists of 0.3-0.6M of trehalose, 0.001-0.004M of polyethylene glycol 8000, 0.02-0.08M of mannitol and the balance of purified water. The multiplex PCR universal freeze-drying protective agent disclosed by the invention can exert effects of aprotective agent and an excipient in a freeze-drying process of a multiplex PCR reagent, and the added protective agent has simple components and does not influence an amplification reaction of the multiplex PCR. After the freeze-drying protective agent is used for freeze-drying, appearance is smooth, hygroscopic performance is low, performance change of the multiplex PCR reagent before and afterfreeze-drying is small, and stability of a freeze-dried product is good. A preferred freeze-drying procedure has high adaptability and can be applied to various glycerol-free enzyme reaction systems under adoption of the freeze-drying protective agent provided by the invention.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a general-purpose freeze-drying protective agent for multiple fluorescent PCR and its application. Background technique [0002] Multiple fluorescent PCR (multiplex PCR), which is to add more than two pairs of primers and probes in the same PCR reaction system, so that two or more different nucleic acid fragments can be amplified and detected by PCR in one reaction system. Quickly obtain the detection results of multiple targets, significantly shortening the detection time. [0003] The freeze-drying of PCR reagents is to add the freeze-drying protection agent to the prepared liquid PCR detection system, freeze it into a solid in advance, and use the sublimation of water under vacuum conditions to dehydrate the materials at low temperature to achieve the purpose of reagent drying. During the freeze-drying process, the heat-sensitive substance (enzyme) will not be denatured or inact...

AI Technical Summary

Problems solved by technology

The main problems are: 1) There is no suitable freeze-drying protective agent, which leads to a decrease in the enzyme activity in the PCR system during the freeze-drying process

2) The added lyoprotectant has a great influence on the multiple amplification performance of PCR reagents, and the overall performance of the reagents decreases after lyophilization

The whole process takes a long time period, heavy workload and high cost

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