A preparation kit of puccinilia selloana cell nucleus suspension and its application method

By employing a lysis buffer, phase separation, and density gradient purification system, the issues of purity and integrity in the extraction of Napier's tail cells were resolved, enabling the efficient and low-cost preparation of cell nucleus suspensions suitable for single-cell omics research.

CN121086969BActive Publication Date: 2026-08-25SICHUAN AGRI UNIV
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Patent Information

Application Number
CN202511233413.9
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2025-09-01
Publication Date
2026-08-25
Estimated Expiration
2045-09-01

AI Technical Summary

Technical Problem

Existing technologies struggle to efficiently extract high-quality nuclei from American foxtail millet cells, especially in the presence of polyphenols and polysaccharides, making it difficult to guarantee the purity and integrity of the nuclei, leading to decreased experimental data quality and increased costs.

Method used

A lysis buffer system, a phase separation system, and a density gradient purification system were employed, including a lysis buffer system composed of Triton X-100, Tween 20, and PVP-40, ATPS two-phase separation buffer, and Percoll separation buffer, combined with specific buffers and staining solutions, to achieve selective lysis, polysaccharide removal, and density gradient separation, ensuring the integrity and purity of cell nuclei.

Benefits of technology

It enables efficient and rapid preparation of high-purity and high-integrity cell nuclear suspensions, suitable for single-cell omics research, improving experimental signal-to-noise ratio and data quality, and reducing experimental costs.

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Abstract

The application provides a preparation kit for a Puccinellia distans cell nucleus suspension and relates to the field of molecular biology, and comprises a lysis buffer system, a phase separation system, a density gradient purification system and a lysis and staining system; wherein the lysis buffer system comprises 1xNIB working solution and 1xTAPS resuspension buffer, the pH value of the 1xTAPS resuspension buffer is 7.8-8.2; the phase separation system adopts ATPS two-phase separation solution; the density gradient purification system adopts 33%-37% Percoll separation solution; and the lysis and staining system comprises a cell lysis solution and a DAPI staining solution. The kit can obtain stable high-quality plant cell nucleus suspension, makes the cell nucleus damage small, improves the nucleus integrity rate, reduces the cell nucleus fragments and reduces the cell nucleus adhesion, and provides key technical support for plant single cell omics research of Puccinellia distans and the like.
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