Small Target Nucleic Acid Isolation Yield Recovery
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Solution Overview
Problem
Current nucleic acid isolation methods, particularly those using column-based solid phases, often result in the loss of small target nucleic acids due to their release during enzymatic and chemical treatments, as restoration buffers are insufficient in re-capturing these nucleic acids, leading to reduced yields.
Innovation Solution
A method involving binding target nucleic acids to a nucleic acid binding solid phase, performing enzymatic and/or chemical treatments, collecting released small target nucleic acids as flow-through, and re-binding them using a recovery solution with a chaotropic agent and alcohol to enhance binding, thereby increasing the yield of small target nucleic acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-generated harmful factors
If enzymatic and chemical treatments are performed on the nucleic acid binding solid phase while target nucleic acids are bound, then protein and contaminant removal is improved, but small target nucleic acids are released and lost during treatment
Solution Approach 1:
The patent extracts small target nucleic acids from the flow-through fraction that contains released nucleic acids, separating them from the main binding process. This allows the treatment steps to proceed while recovering the released small nucleic acids that would otherwise be lost.
Solution Approach 2:
Instead of discarding the flow-through fraction containing released small target nucleic acids, the patent recovers these nucleic acids by applying the flow-through to a fresh nucleic acid binding solid phase, thereby recovering material that would have been lost during the treatment process.
2Stability of the object's composition
If restoration buffer is applied after enzymatic treatment to restore binding conditions, then binding conditions are improved, but small target nucleic acids remain insufficiently re-captured
Solution Approach 1:
The patent uses a fresh nucleic acid binding solid phase as an intermediary to capture small target nucleic acids from the flow-through. This intermediary step effectively transfers the released small nucleic acids to a new binding surface where they can be stabilized and recovered.
3Productivity
If column-based solid phase is used for nucleic acid binding, then isolation efficiency is improved, but small target nucleic acids are released during treatment steps
Solution Approach 1:
The patent segments the isolation process into distinct phases: initial binding to solid phase, treatment steps with flow-through collection, and recovery binding to a second solid phase. This segmentation allows treatment of the bound material while separately capturing released small nucleic acids.
Solution Approach 2:
The patent recovers small target nucleic acids that are released during treatment by applying the flow-through to a fresh nucleic acid binding solid phase, preventing loss of these small molecules while maintaining the efficiency benefits of column-based isolation.
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 significantly enhances the yield of small target nucleic acids by efficiently re-capturing those that would otherwise be lost during the isolation process, improving the overall recovery of these nucleic acids.
Implementation Method 1
contacting said flow-through which comprises small target nucleic acids mixed with a recovery solution with a nucleic acid binding solid phase for binding the contained small target nucleic acids to said nucleic acid binding solid phase
Implementation Method 2
re-binding them using a recovery solution with a chaotropic agent and alcohol to enhance binding
Data Source
AI summary
The present invention pertains to a method for isolating a target nucleic acid including small target nucleic acids from a sample, said method comprising at least the following steps a) binding at least a portion of the target nucleic acid including small target nucleic acids to a nucleic acid binding solid phase comprised in a column by passing the sample through said column, b) performing an enzymatic and/or chemical treatment on the nucleic acid binding solid phase while the target nucleic acid is bound to said solid phase, c) collecting at least a portion of the small target nucleic acids released from the solid phase during said treatment of step b) as flow-through, d) contacting said flow-through which comprises small target nucleic acids mixed with a recovery solution with a nucleic acid binding solid phase for binding the contained small target nucleic acids to said nucleic acid binding solid phase, e) optionally performing an elution. The present invention results in a considerable increase in the yield of small target nucleic acids in the isolated target nucleic acid because it allows to efficiently capture and recover small target nucleic acids.