Nucleic Acid Extraction Reagent for FFPE Tissue
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for nucleic acid extraction from cellular sources, particularly Mycobacteria, yeast, and formalin-fixed paraffin-embedded (FFPE) tissue samples, are complex and multi-step processes, involving multiple solutions and enzymes, which complicate and prolong the extraction process.
Innovation Solution
A one-step method using an aqueous extraction solution comprising polar and non-polar organic solvents, chaotropes, and detergents to solubilize paraffin, break down tissue, and release nucleic acids, with amine monomers like 2,2'-(ethylenedioxy)bis(ethylamine) and chaotropes like urea or guanidine thiocyanate, allowing direct extraction and capture on silica-containing particles without separate de-paraffinization or protease digestion.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple-step extraction methods are used, then extraction reliability is improved, but process complexity and time increase
Solution Approach 1:
The patent combines multiple extraction steps into a single integrated procedure. The extraction solution simultaneously performs deparaffinization, tissue digestion, and nucleic acid extraction in one step, eliminating the need for separate deparaffinization and protease digestion steps while maintaining extraction reliability
Solution Approach 2:
The extraction solution is designed as a universal reagent that performs multiple functions: it solubilizes paraffin, breaks down tissue proteins, and extracts nucleic acids simultaneously. This multi-functional approach simplifies the process while maintaining effectiveness across different sample types
2Reliability
If multiple-step extraction methods are used, then extraction reliability is improved, but processing time increases
Solution Approach 1:
The patent merges deparaffinization, tissue digestion, and nucleic acid extraction into a single simultaneous step, reducing processing time while maintaining reliability through the comprehensive action of the extraction solution
3Reliability
If enzymes are used for tissue breakdown, then extraction effectiveness is improved, but process complexity increases
Solution Approach 1:
The patent extracts and removes the need for enzymatic components by using a chemical extraction solution that directly breaks down tissue proteins and solubilizes paraffin without requiring added enzymes, simplifying the process while maintaining effectiveness
Solution Approach 2:
The extraction solution is designed to perform tissue breakdown and paraffin solubilization intrinsically through its chemical composition, eliminating the need for separate enzymatic treatments and simplifying the overall process
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
This method simplifies the extraction process, eliminating the need for enzymes and multiple steps, achieving efficient and effective extraction of nucleic acids from challenging samples like Mycobacteria and FFPE tissues, with improved DNA yield and reduced processing time.
Implementation Method 1
utilizes a combination of polar and non-polar organic solvents as well as chaotropes and detergents to solubilize the paraffin
Implementation Method 2
utilizes a combination of polar and non-polar organic solvents as well as chaotropes and detergents to solubilize the paraffin, break down the tissue and release the nucleic acids
Implementation Method 3
utilizes a combination of polar and non-polar organic solvents as well as chaotropes and detergents to solubilize the paraffin, break down the tissue and release the nucleic acids
Implementation Method 4
The nucleic acids then can be, for example, captured on silica containing particles in a single solution
Data Source
Figure 1
Figure 2
Figure 3
AI summary
Compositions and methods for the efficient extraction, enrichment and isolation of nucleic acids from fresh, fixed or fixed and embedded cells, tissues, biological materials and cellular source material. Prior art methods for nucleic acid extraction from cellular source materials and, particularly, paraffin embedded tissue samples (e.g., formalin-fixed paraffin-embedded samples: FFPE) involves complicated, multi-step processes.