Acid Zeolite Binding Matrix for DNA-RNA Separation
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Solution Overview
Problem
Current RNA purification methods often result in low yield and purity, especially when isolating RNA from complex biological materials, and are contaminated with DNA due to the structural similarity between DNA and RNA, making it difficult to achieve high-quality RNA samples.
Innovation Solution
The use of a binding matrix that preferentially binds DNA in an acidic dilution buffer or acid zeolites to separate DNA from RNA, allowing for the generation of purified RNA samples with high yield and low DNA contamination, using methods that include contacting the sample with a dilution buffer and a binding matrix, followed by separation to remove DNA-bound particles, thereby isolating pure RNA.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional RNA purification methods (such as Sevag procedure or guanidinium thiocyanate method) are used, then RNA can be isolated from biological sources, but the RNA is contaminated with DNA and other cellular materials, requiring extensive additional manipulation for further purification
Solution Approach 1:
The patent extracts and removes DNA from the RNA sample using a binding matrix that selectively binds DNA. The binding matrix is contacted with the RNA sample, DNA binds to the matrix, and then the matrix is separated from the RNA, leaving purified RNA. This extraction approach directly addresses the contamination issue while maintaining RNA yield.
Solution Approach 2:
The binding matrix serves as an intermediary substance that selectively interacts with DNA. It acts as a mediator between the DNA contamination and the purification process, binding to DNA molecules and enabling their removal from the RNA sample without affecting RNA integrity.
2Adaptability or versatility
If silica based nucleic acid isolation techniques are used, then total RNA can be isolated from simple biological materials, but the yield and purity of RNA is low, particularly when isolating from complex biological materials such as plant or animal tissue
Solution Approach 1:
The patent changes the chemical parameters of the purification system by using an acidic dilution buffer (pH 3-6) in combination with the binding matrix. This parameter change creates conditions that enhance the selective binding of DNA while maintaining RNA stability, thereby improving both purity and yield when isolating from complex biological materials.
3Ease of manufacture
If phenol or phenol-chloroform mixture is used for DNA and RNA isolation, then proteins are denatured and precipitated, but the method is hazardous, laborious, and of limited utility for isolating RNA from sources high in ribonuclease
Solution Approach 1:
The patent converts the harmful effect of RNase into a beneficial outcome. By using the binding matrix to selectively remove DNA, the method eliminates the need for harsh phenol-chloroform extraction that risks RNA degradation. The acidic buffer and binding matrix combination creates conditions where RNase activity is inhibited while DNA is effectively removed, protecting RNA from degradation.
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 achieves high-purity RNA samples with minimal DNA contamination, maintaining at least 75% of the original RNA yield and reducing DNA presence to less than 1% of the total nucleic acid mass, suitable for applications like RT-PCR and diagnostics.
Implementation Method 1
a binding matrix (e.g. a plurality of binding particles or composition coated with a plurality of binding particles) that preferentially binds DNA molecules in the presence of the dilution buffer
Implementation Method 2
a binding matrix that comprises acid zeolites
Implementation Method 3
a binding matrix that comprises acid zeolites
Implementation Method 4
a dilution buffer with an acidic pH
Data Source
AI summary
The present invention relates to methods, kits, and compositions for generating purified RNA samples and purified DNA samples. In particular, the present invention provides methods for generating a purified RNA or DNA sample from a sample containing both DNA and RNA molecules using a binding matrix that preferentially binds DNA or RNA in the presence of an acidic dilution buffer, or using a binding matrix that comprises acid zeolites, as well as compositions and kits for practicing such methods.