Thermoplastic Nanomembrane Size Selection for Nucleic Acids
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Solution Overview
Problem
Current methods for size selection of nucleic acids in sequencing technologies are inefficient, particularly for high molecular weight DNA, as they require separate steps like AMPURE and PFGE, which are slow and damage-prone, and fail to achieve high recovery and purity.
Innovation Solution
The use of nanomembranes, such as NANOBIND, for rapid size selection of nucleic acids by binding, washing, and eluting, which allows for high recovery and purity of DNA across a wide size range without the need for separate purification steps, using a bind, wash, and elute process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If AMPURE beads are used for size selection, then small molecules are removed, but recovery efficiency is low and processing is slow
Solution Approach 1:
The patent replaces the mechanical filtration approach of AMPURE beads with a thermally-driven phase transition system. The nanomembrane utilizes temperature-dependent pore size changes to selectively pass molecules based on size, eliminating the need for bead-based mechanical filtration and enabling faster processing with improved recovery efficiency.
Solution Approach 2:
The patent employs phase transition of the thermoplastic nanomembrane material. By heating the nanomembrane above its glass transition temperature, the pores expand to allow passage of larger molecules. By cooling below this temperature, pores contract to retain smaller molecules. This phase transition mechanism enables rapid, size-selective separation without slow filtration steps.
2Reliability
If separate AMPURE and PFGE purification steps are used, then size selection is achieved, but processing time increases and DNA damage occurs
Solution Approach 1:
The patent merges multiple purification functions into a single nanomembrane device. The same thermoplastic nanomembrane performs both size selection and purification in one integrated step, eliminating the need for separate AMPURE and PFGE steps. This consolidation reduces processing time while maintaining purity through the temperature-controlled selective passage mechanism.
Solution Approach 2:
The nanomembrane is designed with universal applicability for different size selection requirements. By adjusting the temperature to different levels above the glass transition temperature, the same nanomembrane can selectively pass molecules of various sizes, making it a multi-functional tool that replaces multiple specialized purification steps.
3Quantity of substance
If high mass concentration DNA is used for long reads, then molarity is maintained, but recovery of AMPURE and BLUEPIPPIN is impaired
Solution Approach 1:
The patent changes the physical parameter of pore size through temperature control. By heating the nanomembrane above its glass transition temperature, the pores expand to accommodate high mass concentration DNA molecules. This parameter change allows the system to handle high concentrations that would otherwise be rejected by standard low-temperature membranes, thereby improving recovery efficiency.
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 achieves high recovery and purity of nucleic acids, outperforming existing technologies by enabling efficient size selection across a broader size range, reducing processing time, and minimizing DNA damage, thus enhancing sequencing data quality.
Implementation Method 1
a thermoplastic nanomembrane that can rapidly and selectively separate molecules based on size
Implementation Method 2
The thermoplastic polymers can be heated above their glass transition temperature to expand the pores of the nanomembrane
Data Source
AI summary
The present disclosure is directed to a method for purifying a sample containing nucleic acids to obtain isolated nucleic acids of a desired size range, either above a size cut-off, below a cut-off, or within a defined band of sizes, including: a) combining a nucleic acid-containing sample with a binding buffer to provide a binding mixture; b) contacting the binding mixture with a silica nanomembrane, wherein the silica nanomembrane adsorbs nucleic acids from the binding mixture within a desired size-range; and c) separating the bound nucleic acid from the remaining sample. Kits including a silica nanomembrane, a binding buffer and one or wash buffers are also provided herein.