Tumor inhibition factor Let-7a detection kit based on strand displacement and enzyme-assisted circulation signal amplification and use method of tumor inhibition factor Let-7a detection kit

Abstract

The invention belongs to the field of biological analysis, and relates to a tumor inhibition factor Let-7a detection kit based on strand displacement and enzyme-assisted circulation signal amplification. When a target gene exists, through strand displacement reaction, the target gene is assisted to participate in circulation, and a fluorophore in a three-strand composite structure is far away from a quenching group, so that a fluorescence signal is released, and meanwhile, a large amount of A2 single strands are displaced; then the A2 single chain is hybridized with a DNA chain modified with a fluorophore quenching agent in a DNA double-chain track Q + F through a foot point mediated replacement reaction to release an FAM-modified DNA chain with opening fluorescence, and then the DNA chain modified with the fluorophore quenching agent is hybridized with the A2 chain under the action of added excision enzyme, so that the DNA chain modified with the fluorophore quenching agent can be hydrolyzed from the concave end 3 of the DNA chain, the A2 chain is recycled, and a very strong fluorescence signal is generated. The fluorescence signal intensity and the target gene concentration have a certain relationship, so that the sensitive detection of the tumor inhibition factor Let-7a is realized.

Description

technical field [0001] The invention belongs to the field of biological analysis, and relates to a tumor suppressor Let-7a detection kit based on strand displacement and enzyme-assisted circulatory signal amplification and a method for using the same. Background technique [0002] MicroRNA is a non-coding, small fragment (about 20-22 nucleotides in length), single-stranded RNA molecules that play important roles in different physiological processes. Many evidences show that, as a new gene regulator, microRNA can inhibit gene expression. In mammals, microRNA is involved in regulating the activity of more than 60% of protein-coding genes, and the change of microRNA expression is related to the physiological function disorder of the human body, especially are closely related to the tumor process. Therefore, MicroRNAs have been selected as biomarkers in the early diagnosis of many diseases including various cancers. The Let-7 miRNA family has 12 members, of which Let-7a is one...

AI Technical Summary

Problems solved by technology

The patent achieves a low background signal through the recovery of magnetic beads, and under the synergistic action of the shearing enzyme and the polymerase, the signal cycle is amplified, and finally realizes the high-sensitivity detection of the biosensor, but there are still complicated operations and multiple enzymes. The use of the buffer system caused compatibility problems

Images