Low-Conductivity Buffer for DNA Fragment Isolation
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Solution Overview
Problem
Current methods for isolating separated DNA and protein fragments from separation matrices are time-consuming, laborious, and potentially damaging to the DNA, as they involve manual excision and UV illumination, which can nick the DNA.
Innovation Solution
A device and method using a low-conductivity buffer in a sample loading well to improve sample injection and stacking, allowing for precise isolation of separated components within a separation channel, where a low-conductivity buffer is used to confine the electric field and facilitate the separation and collection of specific fragments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If manual excision and UV illumination are used to isolate separated DNA fragments from separation matrix, then DNA fragments can be extracted for further processing, but the process is time-consuming, laborious, and potentially damaging to the DNA
Solution Approach 1:
The patent replaces the manual mechanical excision process with an automated electrophoretic separation and collection system. The device uses electric field-driven migration to transport separated DNA fragments through a separation channel and into collection wells, eliminating the need for manual gel cutting and UV illumination-based extraction, thereby reducing both time and DNA damage
Solution Approach 2:
The system enables self-service by allowing DNA fragments to be automatically separated, transported, and collected based on their electrophoretic migration properties. The device autonomously performs the isolation process without requiring manual intervention for excision and extraction, with separated fragments automatically migrating to collection wells based on size and charge
2Measurement precision
If low-conductivity buffer is used in sample loading well, then sample injection and stacking are improved, but the buffer conductivity must be carefully controlled
Solution Approach 1:
The patent applies local quality by using a low-conductivity buffer specifically in the sample loading well, while the rest of the separation channel uses standard conductivity buffer. This localized buffer with different conductivity properties enables effective sample stacking and injection precision at the loading zone without affecting the overall separation process in the channel
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 efficient and precise isolation of sample components, reducing the risk of DNA damage and improving the resolution and precision of fragment collection, allowing for further analysis such as PCR, ligation, and mass spectrometry.
Implementation Method 1
a low-conductivity buffer is used to confine the electric field
Implementation Method 2
separating the sample material into a plurality of sample components
Data Source
AI summary
The invention provides devices and methods for isolating one or more sample components of a sample material following separation of the sample material into a plurality of sample components. A device includes a separation channel having a sample loading well. A low-conductivity buffer is disposed in the loading well, the buffer having a conductivity<0.2 mS/cm. In a method, a buffer is loaded into a loading well in fluid communication with a separation channel of a device. A sample material having a conductivity higher than that of the buffer is then loaded into the loading well such that the sample material is disposed beneath the buffer, the buffer disposed over and covering the sample material. The sample material is separated into a plurality of separated components in the separation channel, and a separated component is collected from a collection well disposed in a collection leg of the device.