Nucleic Acid Purification via Dual-Stage Binding Matrix
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Solution Overview
Problem
Existing methods for purifying nucleic acids, such as DNA and RNA, from biological samples like blood and tissues often result in low yields and are inefficient when dealing with large samples, especially when trying to extract small amounts of DNA.
Innovation Solution
A method involving a binding matrix and specific formulations to separate and bind nucleic acids, allowing for their purification in a substantially purified form, using conditions that optimize the amount of nucleic acid bound to the matrix based on the surface area, and including the use of agents like guanidine thiocyanate and acetamide derivatives to facilitate separation and binding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If known purification methods are used, then nucleic acid can be extracted from biological samples, but the yield is very low and efficiency is poor when dealing with large samples
Solution Approach 1:
The patent changes the chemical parameters of the binding environment by using guanidine thiocyanate-acetamide buffer instead of traditional buffers. This parameter change creates optimal conditions for nucleic acid binding to the matrix, significantly improving both yield and efficiency of purification from large biological samples
Solution Approach 2:
The patent uses a composite binding matrix consisting of magnetic particles coated with silica and carboxyl groups. This composite structure combines the magnetic properties for easy separation with the chemical properties for specific nucleic acid binding, resolving the contradiction between extraction efficiency and yield
2Quantity of substance
If binding matrix surface area is increased to improve binding capacity, then more nucleic acid can be bound, but the amount bound becomes dependent on surface area rather than being consistent
Solution Approach 1:
The patent changes the chemical composition parameters of the binding buffer to include guanidine thiocyanate and acetamide at specific concentrations. These parameter changes create saturation binding conditions where the chemical environment becomes the limiting factor rather than the surface area, ensuring consistent binding amounts across different matrix quantities
Solution Approach 2:
The binding matrix is designed to function universally across different sample sizes and matrix quantities. The guanidine thiocyanate-acetamide buffer system creates consistent binding conditions regardless of the specific amount of matrix used, making the method universally applicable and reliable
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 efficient purification of nucleic acids, reducing variability in yields and allowing for the extraction of defined quantities, suitable for scientific procedures like PCR, sequencing, and cloning, even from variable biological samples.
Implementation Method 1
combining the medium containing nucleic acid with a definable amount of a binding matrix capable of reversibly binding nucleic acid
Implementation Method 2
The formulations used in the foregoing method can contain guanidine thiocyanate and an amount of (i) acetamide, (ii) one or more acetamide derivatives, or (iii) a combination of acetamide and one or more acetamide derivatives
Implementation Method 3
separating the binding matrix with nucleic acid bound thereto from substantially the rest of the combined medium and formulation
Implementation Method 4
eluting the nucleic acid from the binding matrix, thereby obtaining nucleic acid in a substantially purified form
Data Source
Figure 1
Figure 2
Figure 3A~3B
AI summary
A method and kit which allow the use of a discrete amount of a binding matrix to first purify nucleic acids from a medium under a first set of binding conditions wherein the amount of nucleic acid bound to the binding matrix is essentially independent of the amount of surface area of the definable amount of the binding matrix, followed by a second purification step wherein the nucleic acids are bound to a discrete amount of binding matrix under a second set of binding conditions wherein the amount of nucleic acid bound to the binding matrix is essentially dependent on the amount of surface area of the definable amount of the binding matrix, thus providing a discrete quantity of nucleic acid.