Bead Beating Tube with Dry Blocking Agent for Nucleic Acid Extraction
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Solution Overview
Problem
Current methods for genetic testing and infectious disease diagnosis face limitations due to DNA concentration reduction from sample dilution during cell lysis and nucleic acid extraction, leading to limited detection accuracy and sensitivity.
Innovation Solution
The use of bead beating systems with integrated dry blocking agents, such as urea and detergents, prevents nucleic acid absorption on beads, allowing for higher extraction rates without the need for lysis buffers, thus maintaining sample concentration and enhancing sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If lysis buffer is added to the sample fluid during bead beating, then cell lysis and nucleic acid extraction are facilitated, but the DNA concentration in the sample decreases due to sample dilution
Solution Approach 1:
The patent extracts and removes the lysis buffer step from the traditional bead beating protocol. By performing bead beating directly on the undiluted sample fluid without adding lysis buffer, the method eliminates sample dilution while still achieving effective cell lysis and nucleic acid extraction through the mechanical action of bead beating alone
Solution Approach 2:
The sample fluid itself serves as the extraction medium without requiring external addition of lysis buffer. The bead beating process directly lysed the cells and released nucleic acids into the existing sample fluid, allowing the sample to serve its own extraction function without dilution
2Quantity of substance
If bead beating is performed without blocking agents, then nucleic acid absorption on beads occurs causing loss, but adding blocking agents increases device complexity and may require additional reagents
Solution Approach 1:
The patent employs a disposable bead beating tube pre-coated with blocking agent as an integrated single-use solution. The blocking agent is incorporated into the disposable tube itself rather than requiring separate addition, eliminating the need for complex blocking steps while preventing nucleic acid loss through absorption on beads
Solution Approach 2:
The patent merges the blocking agent function with the bead beating tube structure by pre-coating the tube interior surface with blocking agent. This integration combines the containment function of the tube with the nucleic acid protection function of the blocking agent into a single component, simplifying the overall extraction system
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 enables the recovery of higher quantities of DNA and RNA, improving detection accuracy and sensitivity by preventing nucleic acid loss and maintaining sample concentration during the extraction process.
Implementation Method 1
The beads are adapted to mechanically destroy the cell walls of microorganisms contained in the sample fluid when the sample fluid is filled into the inner cavity and the bead beating tube is subject to mechanical oscillations
Implementation Method 2
the use of bead beating in nucleic acid extraction methods results in nucleic acid loss due to absorption on the beads and that such loss can be prevented by the appropriate amount of blocking agent to block binding sites on the beads
Data Source
AI summary
The present disclosure provides improved methods for bead beating and a bead beating system useful therefor. The bead beating system comprises a sample tube, beads, and a dry blocking agent, and methods for using the bead beating system to extract nucleic acids from cells containing the nucleic acids.