Nucleic Acid Isolation Using Acetic Acid Binding Buffer
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Solution Overview
Problem
Current nucleic acid isolation methods using magnetic particles face challenges in achieving high-concentration nucleic acid extraction efficiently, particularly for molecular diagnosis, due to the need for ethanol removal and the complexity of automation, which can lead to inaccurate results and increased processing time.
Innovation Solution
A method utilizing a binding buffer with a compound having a lower dielectric constant than ethanol, such as acetic acid or phosphoric acid, and a change in pH between buffers to bind nucleic acids to silica magnetic particles, allowing for efficient separation and elution without ethanol, using a magnetic field and basic pH elution buffer.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If ethanol is used in the binding buffer for nucleic acid isolation using magnetic particles, then nucleic acid binding efficiency is improved, but additional processing steps for ethanol removal are required
Solution Approach 1:
The invention extracts ethanol from the binding buffer system and replaces it with acetic acid. This removal of ethanol eliminates the need for subsequent ethanol removal steps while maintaining effective nucleic acid binding to magnetic particles, directly resolving the contradiction between binding efficiency and processing complexity
Solution Approach 2:
The invention changes the chemical parameter of the binding buffer by substituting ethanol with acetic acid. This parameter change maintains the buffer's ability to facilitate nucleic acid binding while eliminating the harmful effect of requiring ethanol removal steps, thus resolving the technical contradiction
2Reliability
If conventional nucleic acid isolation methods are used, then isolation process is established, but processing time is increased due to multiple steps including centrifugation and ethanol removal
Solution Approach 1:
The invention enables continuous processing by eliminating the need for intermediate ethanol removal and centrifugation steps. The acetic acid-based binding buffer allows nucleic acid binding to proceed directly followed by magnetic separation, creating a continuous workflow that reduces processing time while maintaining reliable isolation
Solution Approach 2:
The invention performs the binding function preliminarily using acetic acid in the binding buffer, which prepares the nucleic acid for direct magnetic separation without requiring subsequent ethanol removal. This preliminary action with the modified buffer eliminates later time-consuming steps while maintaining process reliability
3Manufacturing precision
If ethanol is used for washing and binding, then nucleic acid purity is maintained, but automation difficulty increases
Solution Approach 1:
The invention extracts ethanol from the buffer system and replaces it with acetic acid, which does not require special handling or removal steps. This extraction enables easier automation since the acetic acid-based buffers can be handled in standard liquid handling systems without special evaporation or removal protocols, while maintaining nucleic acid purity
Solution Approach 2:
The invention replaces the mechanical complexity of ethanol removal (evaporation, centrifugation) with a simpler chemical system using acetic acid. This substitution allows automated systems to process samples more easily since the acetic acid buffers can be mixed, incubated, and separated using standard magnetic and liquid handling automation without requiring specialized ethanol removal mechanisms
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 enables quick and efficient nucleic acid isolation with improved accuracy and reduced processing time, eliminating the need for ethanol removal and centrifugation, thus facilitating high-concentration nucleic acid extraction suitable for molecular diagnosis.
Implementation Method 1
nucleic acids, the surfaces of which are negatively charged, are allowed to bind to magnetic particles or silica on the surface of the column filter by electrostatic attraction
Implementation Method 2
adding an acidic binding buffer including a compound having a lower dielectric constant than ethanol
Implementation Method 3
separating the magnetic particles by applying a magnetic field
Implementation Method 4
using a lysis buffer, adding an acidic binding buffer including a compound having a lower dielectric constant than ethanol and silica magnetic particles to allow the nucleic acid to bind
Data Source
AI summary
The present invention relates to a method for isolating a nucleic acid by using: a binding buffer containing acetic acid or phosphoric acid; and the pH difference between binding and washing buffers, and an elution buffer, wherein silica magnetic particles are added to a biological sample including a nucleic acid to which a binding buffer containing acetic acid, phosphoric acid, etc., has been added, thereby binding the nucleic acid to the magnetic particles, after which a magnetic field is applied to separate the magnetic particles, and the nucleic acid is purified in a basic pH elution buffer. The method uses the pH difference, and can isolate the nucleic acid more quickly and efficiently than centrifugation or gravity separation. In addition, since ethanol is not used, high-purity nucleic acid can be isolated and purified to obtain more accurate results in subsequent experiments.