Method for detection of amino acids based on aerolysin nanochannel

Abstract

The invention discloses an amino acid detection method based on an aerolysin nanochannel. The method comprises the steps as follows: (1) designing DNA probes connected with formyl groups; (2) connecting amino acid to the DNA probes; (3) preparing the aerolysin bio-nanochannel: I activating aerolysin; II preparing a phospholipid n-decane solution; III preparing a phospholipid bilayer; IV forming the aerolysin bio-nanochannel; (4) detecting amino acid with aerolysin: I acquiring DNA probe single-molecular signals; II acquiring amino acid single-molecular signals; III acquiring DNA probe-amino acid single-molecular signals; IV counting the acquired DNA probe-amino acid single-molecular signals to obtain amino acid characteristic information. Accordingly, different amino acids can be detectedsimultaneously. The method can detect different amino acids simultaneously at the single molecule level, is convenient to operate, real-time and accurate, can realize ultrasensitive resolution and hasvery active significance in life science research.

Description

technical field [0001] The invention relates to a detection technology for analyzing and detecting single molecules in life chemistry, in particular to a method for detecting amino acids based on an aerolysin nanochannel. [0002] technical background [0003] Biological nanochannels are a single-molecule detection method emerging in modern times to detect nucleic acids, polypeptides, and sugars. Currently, the most widely used protein pores are Staphylococcus α-hemolysin (α-hemolsyin) and Mycobacterium smegmatis (MspA). However, due to the limitation of the pore size of the protein pore itself, the above two protein channels are not enough to realize the micro resolution of DNA molecules. The aerolysin biological nanochannel has a smaller pore size, higher sensitivity, and its inner cavity is positively charged, which can effectively slow down the passing rate of DNA, thereby achieving high throughput, label-free, and high sensitivity. , DNA probe-based, single-molecule le...

AI Technical Summary

Problems solved by technology

However, at present, there are not many methods that can simultaneously detect multiple amino acids. Although amino acid analyzers can analyze different amino acids at the same time, the operation is complicated, time-consuming, and relatively high in cost.

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