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Separation and collection device for biological compound

A technology for collecting devices and complexes, applied in biochemical cleaning devices, enzymology/microbiology devices, biological material pretreatment, etc., can solve the problems of low efficiency and cumbersome steps, and achieve the effect of high purity and large quantity

Inactive Publication Date: 2013-07-10
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] After searching the existing literature, it was found that Dapprich, J. & Nicklaus, N. et al. published the article "Connecting DNA to Beads Optically Trapped on Microstructures by Monitoring Bead Displacement" in "Bioimaging" (Dapprich, J. .& Nicklaus, N. DNA attachment to optically trapped beads in microstructures monitored by bead displacement. Bioimaging, 6:25-32, 1998), the paper stated that their research results can obtain a magnetic bead to connect only one DNA fragment, but the The method requires optical tweezers, the steps are cumbersome and the efficiency is low

Method used

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  • Separation and collection device for biological compound
  • Separation and collection device for biological compound

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Such as figure 1 As shown, the separation and collection device of the biological complex includes: a flow chamber 1 with a solution channel inside, and the upstream of the flow chamber 1 solution channel is provided with a sample inlet 2 and an electrophoresis buffer filling port 3, and the electrophoresis The buffer filling port 3 is set as a sheath flow mode; the downstream of the flow chamber 1 solution channel has a purpose complex collection port 6 and an empty micro-nano bead outlet 7, and the purpose complex collection port 6 and the empty micro-nano bead outlet 7 are respectively A collection container is connected; a pair of electrode connection channels are provided on both sides of the downstream of the solution channel of the flow chamber 1, and above the bifurcation of the target complex collection port 6 and the empty micro-nano bead outlet 7, and a positive electrode is arranged at the electrode connection channel. 4 and the negative electrode 5, the pos...

Embodiment 2

[0030] Such as figure 2 As shown, the present embodiment adopts a flow chamber 1 with a solution channel inside, and the position of the positive electrode 4 of the power supply and the collection port 6 of the target compound is located at the same position as the position where the electrophoresis buffer added to the electrophoresis buffer filling port enters the flow chamber channel. The position of the negative electrode 5 of the power supply and the outlet 7 of the empty micro-nano beads are located on the same side as the position when the sample enters the flow chamber. The positive and negative electrodes are connected to a unipolar rectangular pulse power supply.

[0031] Using the above-mentioned device, the product of the connection reaction between fluorescently labeled polymer molecules and micro-nano beads at a molar ratio of 1:100, that is, a single micro-nano bead (purpose complex) connected to a single polymer molecule and empty micro-nano beads The mixture ...

Embodiment 3

[0035] An active group is labeled at one end of the polymer (including DNA, RNA, and a chimera linked to a protein or peptide nucleic acid), and the other end or side chain is labeled with a fluorescent molecule, and after purification, re-use the ligation buffer Dissolution; use another active group-coated micro-nano beads that can react with the active group of the labeled polymer to mix with the above-mentioned treated polymer, and connect the polymer molecules to the micro-nano beads through the linking reaction. beads on. A mixture of single micro-nano beads and empty micro-nano beads connected to a single polymer molecule was obtained, and the surface passivation of the micro-nano beads was the same as in Example 1.

[0036] use as figure 1 The shown flow cell device separates and enriches the complex of a single micro-nano bead carrying a single polymer molecule: add the above-mentioned mixture to the syringe pump connected to the injection port, and add the electropho...

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Abstract

The invention discloses a separation and collection device for a biological compound. The separation and collection device comprises a flow chamber in which a solution channel is formed, wherein a sample injection inlet and an electrophoretic buffer solution filling port are formed at the upstream of the solution channel in the flow chamber; a target compound collection port and a hollow micro nano ball outlet are formed at the downstream of the flow chamber; electrode connection channels are formed in two sides of the solution channel of the flow chamber; a positive and negative electrode pair is arranged at the electrode connection channels; a positive electrode and the target compound collection port are positioned on the same side, and a negative electrode and the hollow micro nano ball outlet are positioned on the same side; and the electrode pair is connected with a continuous direct current power or a single-polarity rectangular pulse power supply. Under the action of an external electric field, a target compound carrying net negative charges enters a target compound collection container, so that the purity of the obtained target compound is high, and the number of the obtained target compound is large; and the separation and collection device can be simply applied to interactive dynamics of single-molecule horizontal research on nucleic acid and nucleic acid as well as nucleic acid and protein, single-molecule nucleic acid sequencing and new-generation sequencing sample preparation.

Description

technical field [0001] The invention relates to a biological compound, in particular to a separation and collection device for a biological compound, especially a separation and collection device for a single polymer molecule carried by a single micro-nano bead. Background technique [0002] Single polymer (DNA, RNA, protein, peptide nucleic acid, etc.) molecules are connected by micro-nano beads for single-molecule manipulation, which can be used to detect the basic single-molecule behavior of various life phenomena, including the interaction between nucleic acid and nucleic acid, nucleic acid and related proteins Action kinetics, antibody screening, nucleic acid single-molecule sequencing and sample preparation for next-generation sequencers, etc. The more common method is to use optical tweezers and magnetic tweezers to manipulate micro-nano beads connected to single-molecule polymers, but how to quickly and easily prepare such single micro-nano beads to connect single po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C07K1/14
Inventor 王志民侯彩玲黄伟东王跃张林霞
Owner SHANGHAI JIAO TONG UNIV
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