Formamide Cleavage Composition for Biotin-Streptavidin Release
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Solution Overview
Problem
Existing methods for cleaving biotin-streptavidin bonds on solid supports are inefficient and often interfere with the hybridization of DNA library fragments to flow cell surfaces, damaging the chemistry on the surface.
Innovation Solution
A biotin-streptavidin cleavage composition comprising 10% to 50% by volume of formamide and a salt buffer is used to cleave the bonds at temperatures below 70°C, allowing for efficient release of DNA library fragments without damaging the flow cell surface chemistry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing methods are used to cleave biotin-streptavidin bonds on solid supports, then bond cleavage is achieved, but the hybridization of DNA library fragments to flow cell surfaces is interfered with and the chemistry on the surface is damaged
Solution Approach 1:
The invention changes the chemical parameters of the cleavage composition by incorporating formamide (10-50% v/v) combined with salt buffer, which modifies the cleavage mechanism to occur at lower temperatures (below 70°C) and prevents damage to the flow cell surface chemistry while maintaining effective biotin-streptavidin bond cleavage
Solution Approach 2:
The formamide-salt buffer composition acts as an intermediary agent that mediates the cleavage process, enabling bond breakage through a gentler chemical mechanism that does not interfere with DNA hybridization or damage the flow cell surface chemistry
2Reliability
If existing methods are used to cleave biotin-streptavidin bonds, then bond cleavage is achieved, but the hybridization efficiency is reduced
Solution Approach 1:
By changing the temperature parameter to below 70°C and adjusting the chemical composition to include formamide and salt buffer, the invention maintains high hybridization efficiency while achieving effective bond cleavage, thus improving productivity
3Reliability
If high temperature is used to cleave biotin-streptavidin bonds, then bond cleavage is achieved, but the flow cell surface chemistry is damaged
Solution Approach 1:
The invention changes the temperature parameter from high temperature to below 70°C by using formamide-salt buffer composition, which enables effective bond cleavage at lower temperatures without damaging the flow cell surface chemistry
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
The cleavage composition effectively breaks biotin-streptavidin bonds at lower temperatures, ensuring rapid hybridization of DNA library fragments to the flow cell surface without affecting the chemistry, and can be used in a dual release mechanism for controlled spatial release.
Implementation Method 1
A biotin-streptavidin cleavage composition comprising from 10% to 50% by volume of formamide and a salt buffer is used to cleave the bonds at temperatures below 70°C
Implementation Method 2
A biotin-streptavidin cleavage composition comprising from 10% to 50% by volume of formamide and a salt buffer
Data Source
Figure 1A~1B
Figure 2A~2B
Figure 2C
AI summary
An example of a biotin-streptavidin cleavage composition includes a formamide reagent and a salt buffer. The formamide reagent is present in the biotin-streptavidin cleavage composition in an amount ranging from about 10% to about 50%, based on a total volume of the biotin-streptavidin cleavage composition. The salt buffer makes up the balance of the biotin-streptavidin cleavage composition. In some examples, the biotin-streptavidin cleavage composition is used to cleave library fragments from a solid support. In other examples, other mechanisms are used to cleave library fragments from a solid support.