Preparation method and application of Asian cotton root tip paraffin section

By applying RNA in situ hybridization methods in Asian cotton root tips, combined with paraffin section technology and single-cell transcriptome sequencing, the problems of high sensitivity and high specificity of cotton root tip gene positioning were solved, and the visual positioning of cotton root tip genes and accurate positioning of tissue site markers were achieved.

CN120651607APending Publication Date: 2025-09-16ANYANG INST OF TECH +1
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Patent Information

Application Number
CN202510740786.9
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2022-07-29
Publication Date
2025-09-16

AI Technical Summary

Technical Problem

Existing RNA in situ hybridization technology has not yet been applied to Asian cotton root tips, making it difficult to achieve high sensitivity and high specificity in locating specific genes. In addition, the polysaccharide and polyphenol components of cotton root tips affect the hybridization effect.

Method used

Provided is an RNA in situ hybridization method, comprising preparing sense and antisense probes, processing cotton root tips by paraffin section technology, combining single-cell transcriptome sequencing, and adopting pre-hybridization, hybridization incubation and color development steps to ensure the accuracy and visualization of gene positioning.

Benefits of technology

High-sensitivity and high-specificity positioning of cotton root tip genes was achieved, the influence of polysaccharides and polyphenols was avoided, and structurally intact paraffin sections were prepared to support the application of single-cell transcriptome sequencing technology.

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Abstract

The invention discloses a preparation method and application of an Asian cotton root tip paraffin section. The preparation method comprises the following steps: S21, culturing an experimental material; s22, taking and fixing experimental materials; s23, dehydrating the experimental material; s24, transparency of the experimental material; s25, carrying out wax dipping on the experimental material; s26, embedding the experimental material; s27, performing wax repairing on the experimental material; s28, preparing hybridized slices of experimental materials; the prepared section can be used for carrying out positioning research on the Asian cotton root tip gene by adopting an RNA in-situ hybridization method, the method has very high sensitivity and specificity, and the obtained hybridization section result can be stored for a long time.
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