Clonal Substrate Enrichment with Barcode Capture and Magnetic Beads
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing sequencing techniques face issues with the presence of polyclonal supports, which reduce the number of usable reads and hinder the efficiency of sequencing processes due to the blending of monoclonal and polyclonal supports.
Innovation Solution
A method is employed to enrich clonal populations of nucleic acids by using supports with enrichment barcodes, where monoclonal supports are isolated using capture probes and magnetic beads, allowing for the separation of target nucleic acids with complementary barcode sequences, thereby increasing the concentration of monoclonal supports relative to polyclonal ones.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If sequencing techniques use supports with multiple copies of target sequence, then sequencing can be performed, but polyclonal supports blend with monoclonal supports reducing the number of usable reads
Solution Approach 1:
The patent segments the support population into monoclonal and polyclonal subsets by introducing unique barcodes to each monoclonal support. This segmentation allows subsequent separation based on barcode sequences, enabling selective retention of monoclonal supports and removal of polyclonal supports, thereby resolving the blending issue while maintaining sequencing productivity
Solution Approach 2:
The patent extracts and removes polyclonal supports from the mixed population through selective capture. By using capture probes that specifically bind to barcode sequences present only on monoclonal supports, the method extracts and isolates monoclonal supports while leaving polyclonal supports behind, thus purifying the support population and increasing the number of usable reads
2Productivity
If polyclonal supports are present in the population, then supports can be generated, but sequencing efficiency is reduced
Solution Approach 1:
The patent applies local quality by assigning unique barcode sequences to specific monoclonal supports, creating distinguishable local identifiers within the support population. This allows selective targeting and enrichment of monoclonal supports based on their unique barcode characteristics, thereby increasing their concentration and improving sequencing efficiency
Solution Approach 2:
The patent implements feedback through the barcode sequencing and analysis process. After initial sequencing, the barcode sequences are analyzed to identify monoclonal supports, and this information feeds back into the selection process where capture probes are designed based on the identified barcode sequences, enabling iterative enrichment of monoclonal supports and continuous improvement of sequencing efficiency
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 enhances sequencing performance by increasing the proportion of monoclonal supports, leading to improved analysis techniques and more effective data acquisition during sequencing.
Implementation Method 1
magnetic beads, allowing for the separation of target nucleic acids with complementary barcode sequences
Data Source
AI summary
A method for enriching clonal populations includes, for a population of target nucleic acids, includes exposing the population of target nucleic acids to a plurality of supports; amplifying the bound target nucleic acids in the presence of a primer to form supports including a plurality of copies of the target nucleic acids of the population of target nucleic acids; applying to the supports a capture primer; applying a magnetic bead functionalize with a moiety to bind to the binder moiety; and separating the first set of supports from the second and third set of supports.