Plant Nucleic Acid Isolation Using Asymmetric Particle Lysis
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Solution Overview
Problem
Nucleic acid isolation from plants is challenging due to the difficult-to-lyse nature of plant cells and the presence of inhibitors, leading to low yields and poor quality of nucleic acids, particularly DNA, and existing methods are not effective across a variety of plant types.
Innovation Solution
A method involving mechanical disruption with non-spherical solid disrupting particles and a chaotropic agent, followed by protein precipitation and inhibitor removal, to isolate nucleic acids efficiently from various plant samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If standard mechanical disruption methods (mortar and pestle or spherical beads) are used, then lysis is achieved, but nucleic acid yield is low and inhibitors remain
Solution Approach 1:
The patent employs non-spherical solid disrupting particles with asymmetric geometries (cubes, triangles, irregular shapes) instead of conventional spherical beads. These asymmetric particles create more effective mechanical disruption forces that efficiently break open plant cell walls while minimizing the release of inhibitors, thereby increasing nucleic acid yield and purity simultaneously
Solution Approach 2:
The patent modifies the physical parameters of the disrupting particles by changing their shape from spherical to non-spherical forms. This parameter change in particle geometry fundamentally alters the mechanical disruption mechanism, enabling more effective cell lysis with reduced inhibitor co-release, thus resolving the contradiction between yield and inhibitor presence
2Productivity
If mortar and pestle is used for lysis, then plant material is disrupted, but the process is time-consuming and difficult to use with multiple samples
Solution Approach 1:
The patent replaces the manual mortar and pestle mechanical system with an automated bead beating system using non-spherical particles. This substitution transforms a labor-intensive, time-consuming manual process into an automated, high-throughput mechanical disruption method that can process multiple samples simultaneously with minimal operational complexity
Solution Approach 2:
The patent employs periodic mechanical disruption cycles using the bead beating system with non-spherical particles. The periodic agitation and disruption cycles enable efficient processing of multiple samples in parallel, dramatically increasing productivity while maintaining ease of operation through standardized protocols
3Adaptability or versatility
If commercially available kits with spherical ceramic or metal beads are used, then lysis is achieved, but the method is non-standard and cannot be applied to multiple different plant types with similar success
Solution Approach 1:
The patent develops a universal protocol using non-spherical solid disrupting particles that can be applied across diverse plant types (herbaceous, woody, aquatic, terrestrial) with consistent effectiveness. The asymmetric particle geometry provides a multi-functional disruption mechanism that adapts to different plant cell wall structures, achieving both high versatility and high nucleic acid yield across all plant types
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method achieves high-quality nucleic acid isolation with high yield, effectively removing inhibitors and improving lysis efficiency across different plant types, resulting in a universal protocol for diverse plant samples.
Implementation Method 1
lysing a plant sample by mechanically disrupting the plant sample in a liquid lysis composition which comprises at least one chaotropic agent
Implementation Method 2
mechanically disrupting the plant sample in a liquid lysis composition which comprises at least one chaotropic agent and one or more solid disrupting particles
Implementation Method 3
contacting the lysed sample with at least one protein precipitating agent
Implementation Method 4
contacting the lysed sample with at least one protein precipitating agent and at least one inhibitor removing agent
Data Source
AI summary
Provided is a method for isolating nucleic acids from a plant sample comprising (a) preparing a lysed sample wherein preparing comprises (i) lysing a plant sample by mechanically disrupting the plant sample in a lysis solution which comprises at least one chaotropic agent and one or more solid disrupting particles, and (ii) optionally clearing the lysate; (b) contacting the lysed sample with at least one protein precipitating agent and at least one inhibitor removing agent and providing a mixture; (c) obtaining a liquid phase from the mixture; and (d) isolating nucleic acids from the liquid phase. Also provided is a kit for use in such method.