Universal Lysis Buffer for Plant DNA Extraction
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Solution Overview
Problem
Conventional lysis buffers for DNA extraction from plants are costly, inefficient, and not universally applicable across various plant species and types of plant material, often requiring multiple steps and resulting in residual impurities that inhibit downstream applications.
Innovation Solution
A novel lysis buffer composition comprising Tris, HEPES, MOPS, EDTA, SDS, and glycerol, which provides a universal solution for DNA extraction from a wide variety of crops and plant materials, reducing the need for filtration and ethanol precipitation steps, thereby lowering costs and increasing DNA yield and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional lysis buffers and commercial kits are used for DNA extraction from plants, then DNA can be recovered, but the cost per sample analyzed increases and residual impurities inhibit downstream applications
Solution Approach 1:
The patent modifies the chemical composition parameters of the lysis buffer by replacing conventional components (SDS, EDTA, Tris) with alternative compounds including beta-mercaptoethanol, polyvinylpyrrolidone, and specific salt concentrations. These parameter changes eliminate the need for Proteinase K enzyme and filtration steps, reducing costs while maintaining DNA purity suitable for downstream applications like PCR and sequencing.
Solution Approach 2:
The patent extracts and removes unnecessary processing steps from the conventional DNA extraction protocol. By formulating a lysis buffer that achieves complete cell lysis and DNA release without requiring filtration or additional purification steps, the method eliminates these intermediate processes, thereby reducing cost and time while maintaining DNA quality.
2Reliability
If multiple processing steps including filtration are used in conventional methods, then DNA purity is improved, but the complexity and cost per sample increases
Solution Approach 1:
The patent merges multiple functions into the single lysis buffer formulation. The buffer simultaneously performs cell wall disruption, cell membrane lysis, protein denaturation, and DNA stabilization without requiring separate filtration or purification steps. This consolidation reduces device complexity while maintaining DNA purity through the synergistic action of buffer components.
Solution Approach 2:
The lysis buffer is designed as a universal solution that works across diverse plant species and tissue types without requiring method modification. The multi-functional buffer handles various plant cell wall compositions and intracellular environments, eliminating the need for species-specific optimization and multiple processing steps while ensuring consistent DNA purity outcomes.
3Adaptability or versatility
If conventional lysis buffers are used, then DNA extraction can be performed, but the buffers are not universally applicable across various plant species and types of plant material
Solution Approach 1:
The patent develops a universal lysis buffer formulation that effectively lysed cells from diverse plant species including corn, soy, wheat, and various vegetative tissues. The buffer achieves this universality through its composition of beta-mercaptoethanol for disulfide bond disruption, polyvinylpyrrolidone for polysaccharide binding, and optimized salt concentrations that work across different plant cell wall compositions, thereby maintaining reliable DNA extraction efficiency across all plant types without requiring method modification.
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 novel lysis buffer enables efficient, cost-effective DNA extraction with high yields and purity, suitable for automated methods and various analysis techniques, including SNP studies and marker-assisted breeding, without the need for filtration or extensive clean-up, making it suitable for both manual and automated protocols.
Implementation Method 1
disruption of plant tissues and cells, DNA release into the extraction buffer
Implementation Method 2
sodium dodecyl sulfate (SDS)
Implementation Method 3
ethylenediaminetetraacetic acid (EDTA)
Data Source
AI summary
The present invention is generally directed to a lysis buffer for extraction of DNA from plant material and improved methods for extraction of DNA from plant material utilizing the novel lysis buffer. Advantageously, the lysis buffer of the present invention is suitable for use in connection with simpler analysis methods, while still providing suitable DNA yields and purities for analysis.