Hydrogel Flow Cell Surface Regeneration for Reusable Sequencing
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Solution Overview
Problem
Existing flow cells used in nucleic acid analysis face challenges in maintaining surface chemistry integrity throughout multiple sequencing cycles, as the functional groups for primer attachment degrade after initial use, necessitating the development of a method to regenerate these groups for repeated use.
Innovation Solution
The flow cells incorporate a polymeric hydrogel with functional groups that can attach primers, and after sequencing, reactions are performed to remove the primers and regenerate these functional groups, enabling subsequent cycles of primer grafting and sequencing through processes like cross-metathesis or conversion of post-sequencing groups back to primer-attaching capabilities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If covalent linking is used to maintain surface chemistry in the active area, then reliability is improved, but the flow cell cannot be reused for multiple sequencing cycles
Solution Approach 1:
The patent implements a primer removal process that discards the used primers from the flow cell surface through chemical treatment, and then recovers the flow cell by regenerating the functional groups to their original state, enabling multiple sequencing cycles
Solution Approach 2:
The patent changes the chemical state of the functional groups through controlled reactions - converting them from primer-bound states back to primer-ready states through chemical treatment, thereby extending the flow cell's operational lifespan
2Duration of action of stationary object
If primers are removed after sequencing, then the flow cell can be reused, but the functional groups become different and lose primer attachment capability
Solution Approach 1:
The patent applies chemical treatment to change the chemical parameters of the functional groups, converting post-sequencing functional groups back into the original functional groups capable of attaching primers, thus restoring primer attachment capability
3Duration of action of stationary object
If cross-metathesis reaction is used for primer removal, then functional groups are regenerated, but additional chemical processing is required
Solution Approach 1:
The patent uses cross-metathesis reaction as an intermediary chemical process that temporarily modifies the functional groups during primer removal, then restores them to their original state, enabling flow cell reuse despite the added processing step
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 the reusability of flow cells, maintaining their functionality over multiple sequencing cycles, reducing the need for consumable replacement and enhancing the efficiency and cost-effectiveness of nucleic acid sequencing processes.
Implementation Method 1
the surface chemistry is covalently bound to the flow cell surface
Implementation Method 2
primer removal involves a cross-metathesis reaction that introduces the functional groups that are capable of attaching the primers
Implementation Method 3
the post-sequencing functional groups are converted back into the functional groups that are capable of attaching the primers
Data Source
AI summary
An example of a kit includes a flow cell, a primer fluid, and a cleaving fluid. The flow cell includes at least one surface functionalized with a polymeric hydrogel including azide functional groups or amine functional groups. The primer fluid includes a plurality of alkyne-containing primers, each alkyne-containing primer having an amino cleavable group attaching a primer sequence of the alkyne-containing primer to an alkyne-containing moiety of the alkyne-containing primer. The cleaving fluid includes a substance that is reactive with the amino cleavable group.