Nucleic Acid Orientation Identification via Barcode Partitioning
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for processing biological samples, such as PCR and sequencing, lack efficient means to identify the sequence and orientation of nucleic acid molecules, particularly in separating and analyzing nucleic acid molecules with mutations for accurate sequencing and orientation identification.
Innovation Solution
The method involves subjecting nucleic acid molecules to conditions that introduce mutations, partitioning them with barcode molecules in discrete environments, and generating barcoded nucleic acid molecules for sequencing, which allows for the identification of sequence and orientation information by aligning sequences with reference genomes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If nucleic acid molecules are subjected to conditions sufficient to provide mutations and processed in discrete partitions with barcode molecules, then orientation information and sequence identification are improved, but device complexity and process complexity increase
Solution Approach 1:
The nucleic acid sample is divided into discrete partitions (droplets or wells), each containing individual nucleic acid molecules and barcode molecules. This segmentation enables independent processing of each molecule, allowing accurate sequence identification and orientation determination without interference from other molecules, thus resolving the contradiction between measurement precision and process complexity by organizing the complex process into manageable discrete units
Solution Approach 2:
Barcode molecules are pre-loaded into discrete partitions before nucleic acid molecules are introduced. This preliminary action ensures that when nucleic acid molecules undergo mutation and processing, the barcodes are already in position to capture and record orientation information, simplifying the overall process while maintaining high measurement precision
2Loss of information
If discrete partitions with barcode molecules are used for nucleic acid processing, then orientation information is obtained, but manufacturing complexity increases
Solution Approach 1:
The barcode molecules within discrete partitions self-associate with nucleic acid molecules through complementary base pairing. This self-service mechanism automatically captures orientation information without requiring external intervention or complex manufacturing steps, thus retaining orientation information while maintaining ease of manufacture
Solution Approach 2:
Barcode molecules act as intermediaries between the nucleic acid molecules and the detection system. They capture orientation information through specific binding interactions and transfer this information in a standardized format that can be easily read and analyzed, thus preserving orientation information while simplifying the manufacturing and detection process
Data Source
AI summary
The present disclosure provides methods and systems for nucleic acid preparation and analysis. Nucleic acid molecules may be derived from one or more cells. Preparation of nucleic acid molecules may comprise generation of one or more mutations, such as strand-specific mutations. Nucleic acid molecules may be prepared for and analyzed by sequencing. Sequences may be identified with nucleic acid orientation information.