Universal Primer Concurrent Nucleic Acid Detection
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Solution Overview
Problem
Current nucleic acid sample preparation methods are inefficient and limited, as they often require physical separation of RNA and DNA, making it difficult to detect both forms simultaneously, especially in samples with low quantities or poor quality nucleic acids, leading to potential loss of valuable information.
Innovation Solution
The development of methods and systems that enable concurrent processing and detection of multiple nucleic acid forms, such as single-stranded DNA, double-stranded DNA, single-stranded RNA, and double-stranded RNA, within a single reaction mixture without the need for physical separation, using adapters and specific enzymes like CircLigase II and Bst 2.0 DNA polymerase to tag and amplify these forms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If physical separation of RNA and DNA is performed, then detection of each nucleic acid form can be achieved, but sample processing efficiency decreases and valuable information may be lost due to insufficient material
Solution Approach 1:
The patent combines the detection of RNA and DNA into a single reaction mixture by using a universal primer that can bind to both RNA and DNA templates. The method merges previously separate processing workflows into one concurrent process, allowing simultaneous amplification and detection of both nucleic acid forms without physical separation, thereby maintaining sample integrity and processing efficiency while achieving accurate detection of each form
2Adaptability or versatility
If sample is divided for parallel processing of RNA and DNA, then both forms can be detected, but quantity of nucleic acid material decreases leading to potential loss of valuable information
Solution Approach 1:
The patent employs a universal primer with multi-functionality that can serve as a primer for both RNA and DNA templates. This single primer performs multiple functions: it binds to RNA templates for reverse transcription, binds to DNA templates for direct amplification, and enables concurrent detection of both nucleic acid forms. The universal primer eliminates the need to divide the sample into separate portions, allowing full utilization of the available nucleic acid material while maintaining versatility in detecting both RNA and DNA
3Ease of manufacture
If current sample preparation methods are used, then processing can be performed, but the methods are inefficient and have limited uses
Solution Approach 1:
The patent performs preliminary action by designing a universal primer that is pre-configured to recognize and bind to both RNA and DNA templates. This pre-designed primer eliminates the need for separate primer preparation and sample division steps. The method simplifies the overall workflow by incorporating the differentiation capability into the primer design itself, allowing direct concurrent processing of both nucleic acid forms in a single reaction mixture without complex sample preparation procedures
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 allows for more efficient and accurate genetic analysis by enabling the simultaneous detection of multiple nucleic acid forms, improving the analysis of genetic material in samples with low quantities or poor quality nucleic acids, and providing comprehensive genetic information.
Implementation Method 1
using adapters and specific enzymes like CircLigase II and Bst 2.0 DNA polymerase to tag and amplify these forms
Implementation Method 2
using adapters and specific enzymes like CircLigase II and Bst 2.0 DNA polymerase to tag and amplify these forms
Implementation Method 3
annealing a first primer to the first adapter and annealing a second primer to the second adapter
Data Source
AI summary
The disclosure provides methods, compositions, systems, and kits for the concurrent detection and analysis of different structural and chemical forms of nucleic acids in a sample.