Nucleic Acid Purification via Guanidine Thiocyanate-Acetamide Formulation

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Solution Overview

Problem

Existing methods for purifying nucleic acids from biological samples, such as human blood and tissues, often result in low yields and are inefficient when dealing with small amounts of nucleic acids from large samples.

Innovation Solution

A method using a formulation containing guanidine thiocyanate and acetamide derivatives to separate nucleic acids from their cellular environment, allowing them to bind to a binding matrix, followed by elution for purification, which can also be used to purify multiple nucleic acids simultaneously.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If known purification methods are used, then nucleic acids can be separated from biological samples, but the yield is very low and efficiency is poor when dealing with small amounts of nucleic acids from large samples

Engineering Contradiction:
Improvepurification efficiencyVSAvoidnucleic acid yield
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The invention changes the chemical parameters of the purification system by introducing guanidine thiocyanate-acetamide formulation and adjusting pH to specific ranges (pH 3-6 for DNA, pH 7-10 for RNA), which fundamentally alters the binding characteristics and enables high-yield purification

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention uses composite binding matrices such as silica-cellobiose and silica-carboxy-cellulose, which combine different materials to achieve both high binding capacity and selectivity for nucleic acids, resolving the contradiction between efficiency and yield

Inventive Principle:
Principle #40Composite materials

2Loss of time

If conventional purification methods are applied, then some nucleic acid separation is achieved, but the process is time-consuming and requires multiple steps

Engineering Contradiction:
Improvepurification timeVSAvoidpurification throughput
Core Design Contradiction:
Loss of timeVSProductivity

Solution Approach 1:

The invention merges cell lysis, nucleic acid binding, and purification into a single integrated step by adding formulation directly to biological samples and incubating at room temperature, eliminating the need for separate lysis and purification steps

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The binding matrices are pre-modified with specific chemical groups (cellobiose, carboxy-cellulose) that are pre-conditioned to bind nucleic acids under physiological conditions, eliminating the need for additional activation or preparation steps during the purification process

Inventive Principle:
Principle #10Preliminary action

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 substantial purification of nucleic acids, including DNA and RNA, from various samples, even those with inhibitors, and allows for the storage and transport of bound nucleic acids for later use in scientific procedures.

Implementation Method 1

the respective amounts of GTC and acetamide and/or acetamide derivative(s) present in the formulation are sufficient to cause the at least one nucleic acid to separate from its in vivo cellular environment and bind to the binding matrix

Methodology Applied
Scientific EffectChemical separation:

Implementation Method 2

bind to the binding matrix

Methodology Applied
Scientific EffectAdsorption: Adsorption

Data Source

PatentEP2592143B1Methods of purifying a nucleic acid and formulation and kit for use in performing such methods
Publication Date: 2015.10.07 PROMEGA CORP
  • EP2592143B1 patent drawingFigure 1
  • EP2592143B1 patent drawingFigure 2
  • EP2592143B1 patent drawingFigure 3A~3B

AI summary

A formulation containing guanidine thiocyanate together with acetamide, one or more acetamide derivatives, or a combination of acetamide and one or more acetamide derivatives is used to purify one or more nucleic acids contained in a medium. In particular, a medium containing at least one nucleic acid is combined with a binding matrix and the formulation in order to cause the at least one nucleic acid to separate from its in vivo cellular environment and to bind to the binding matrix. The binding matrix with at least one nucleic acid bound thereto then is separated from substantially the rest of the combined medium and formulation, after which the at least one nucleic acid is eluted from the binding matrix to obtain the at least one nucleic acid in a substantially purified form. If different nucleic acids are to be selectively purified from a single medium, multiple binding matrices, each compatible with a different nucleic acid, can be used.