Compositions and methods for preserving DNA methylation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
DNA methylation patterns are prone to deviation from original patterns when stored at room temperature for extended periods, necessitating improved storage conditions to preserve these patterns for prolonged times.
Innovation Solution
A composition comprising a biological sample with a methylated DNA sequence in a storage solution containing methanol and tris(hydroxymethyl)aminomethane (tris) at specific pH and concentration, optionally with butylated hydroxytoluene (BHT), effectively preserves methylation patterns.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If biological samples are stored at room temperature for extended periods, then storage convenience and accessibility are improved, but DNA methylation patterns deviate from original patterns
Solution Approach 1:
The patent applies parameter changes by modifying the chemical composition of the storage solution, specifically incorporating methanol at concentrations of 50-100% and Tris buffer at pH 7.5-8.5. These parameter changes in the storage medium enable room temperature storage while preserving DNA methylation patterns, thus resolving the contradiction between storage convenience and methylation fidelity.
2Reliability
If biological samples are stored at low temperatures (e.g., -80°C), then DNA methylation patterns are preserved, but storage cost and operational complexity increase
Solution Approach 1:
The patent employs a disposable storage solution formulation that allows samples to be stored at room temperature without requiring expensive cryogenic infrastructure. The methanol-Tris-based solution acts as a stable, long-term preservative that eliminates the need for continuous cold chain maintenance, thereby reducing device complexity and operational costs while maintaining methylation pattern integrity.
3Ease of operation
If standard storage solutions (e.g., PBS, TE buffer) are used, then sample accessibility is maintained, but methylation patterns are not preserved over time
Solution Approach 1:
The patent utilizes a composite storage solution formulation combining methanol (50-100%), Tris buffer (pH 7.5-8.5), and optionally BHT (1-100 ppm). This composite material leverages the preservative properties of methanol, the pH stabilization of Tris, and the antioxidant protection of BHT to create a synergistic system that maintains both sample accessibility and methylation pattern stability, overcoming the limitations of standard single-component buffers.
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 methylation patterns are maintained with high fidelity, retaining at least 60-95% similarity compared to pre-storage levels for up to two years at various temperatures, including room temperature.
Implementation Method 1
a storage solution comprising methanol and tris(hydroxymethyl)aminomethane (tris); wherein methylation patterns of the methylated DNA sequence are preserved
Implementation Method 2
the storage solution further comprises butylated hydroxytoluene (BHT)
Data Source
AI summary
The disclosure provides methods for storage solutions for preserving DNA methylation patterns over a period of time. The disclosure also provides for methods of using methylated DNA stored in such storage solutions.