Spin-Column Nucleic Acid Isolation from Microvesicles
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for isolating nucleic acids from biological samples, particularly cell-free DNA and microvesicles, are slow, tedious, and not scalable, leading to variability in results and inefficiencies in nucleic acid extraction.
Innovation Solution
The method involves capturing DNA and microvesicles onto a surface, lysing them to release nucleic acids, and using a spin-column based purification process with a phenol-based reagent and silica column to isolate and extract DNA and RNA, allowing for efficient and scalable extraction of high-quality nucleic acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ultracentrifugation is used to isolate and extract nucleic acids from biological samples, then nucleic acids can be isolated from microvesicles, but the process becomes slow, tedious, and not scalable
Solution Approach 1:
The patent replaces the mechanical ultracentrifugation system with a chemical-based spin column purification system. Instead of using high-speed centrifugal force to separate microvesicles and nucleic acids, the invention uses silica membrane columns with specific chemical properties that selectively bind nucleic acids under controlled buffer conditions, enabling faster and more scalable processing
Solution Approach 2:
The patent changes the physical and chemical parameters of the isolation process by using spin columns with specific pore sizes, silica surface properties, and buffer compositions. These parameter changes allow nucleic acid binding at controlled pH and salt concentrations, followed by selective elution, replacing the time-consuming ultracentrifugation process with a rapid column-based method
2Reliability
If ultracentrifugation is used to isolate microvesicles, then microvesicles can be separated from supernatant, but the process requires multiple steps including washing, lysing, and column purification
Solution Approach 1:
The patent combines multiple separate processing steps into a single integrated spin column purification process. Instead of performing ultracentrifugation, washing, lysis, and column purification as separate operations, the invention integrates these functions into one device where the silica membrane column performs selective binding, washing, and elution in sequence, reducing the number of manual操作步骤
Solution Approach 2:
The spin column device is designed to perform multiple functions: it acts as a filter to retain microvesicles, a binding matrix to capture nucleic acids, a washing system to remove contaminants, and an elution device to release purified nucleic acids. This multi-functional design eliminates the need for separate equipment for each step
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 approach results in fast, robust, and scalable isolation of high-quality nucleic acids, improving the accuracy and sensitivity of diagnostic applications by enabling the detection of low-frequency sequence variants and reducing the need for invasive tissue biopsies.
Implementation Method 1
the capture surface is an anion exchanger... the anion exchanger is functionalized with quaternary ammonium
Implementation Method 2
a lysis reagent is used to perform on-membrane lysis and release of the nucleic acids
Implementation Method 3
Chloroform extraction is then performed using PLG tubes
Implementation Method 4
The nucleic acids are then bound to a silica column, washed and eluted
Data Source
Figure 1
Figure 2
Figure 3
AI summary
The invention provides novel methods and kits for isolating nucleic acids from biological samples, including cell-free DNA and/or cell-free DNA and nucleic acids including at least RNA from microvesicles, and for extracting nucleic acids from the microvesicles and/or from the biological sample