Electromagnetic method and device for controlling single-chain nucleic acid perforating speed

A single-stranded nucleic acid, perforation speed technology, applied in the field of bioengineering, can solve problems that cannot be used in actual sequencing

Inactive Publication Date: 2007-06-27
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, nanopore single-molecule sequencing is still in the critical stage and cannot be used for actual sequencing.

Method used

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  • Electromagnetic method and device for controlling single-chain nucleic acid perforating speed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Control of perforation speed of BAC single-stranded DNA

[0016] ①The complex connecting a single-stranded target DNA molecule and a magnetic bead is fixed on the negative pole of the electrophoresis tank with an electromagnet; the two poles of the electrophoresis tank are separated by nanopores.

[0017] ②Turn on the power supply, and give a pulling force (540pN) on the free end of the negatively charged target molecule to the positive pole. Since the end of the target molecule connected to the magnetic bead is fixed by the electromagnet (the electromagnetic force is 50pN), the result is that the target molecule is straightened. Then through fine-tuning, the electromagnetic force is weakened and the magnetic beads are released, but the electromagnet still attracts the magnetic beads, so that the target molecule moves slowly to the positive pole, and the speed of passing through the nanopore is controlled at 0.5 bases per millisecond, and the patch clamp To r...

Embodiment 2

[0019] Example 2: Control of the perforation speed of mRNA

[0020] ①The complex connecting a single-stranded target RNA molecule and a magnetic bead is fixed with an electromagnet on the negative pole of the electrophoresis tank; the two poles of the electrophoresis tank are separated by nanopores.

[0021] ②Turn on the power supply, and give a pulling force (1000pN) to the free end of the negatively charged target molecule towards the positive pole. Since the end of the target molecule connected to the magnetic bead is fixed by the electromagnet (the electromagnetic force is 1000pN), the result is that the target molecule is straightened. Then through fine-tuning, the electromagnetic force is weakened and the magnetic beads are released, but the electromagnet still attracts the magnetic beads, so that the target molecule moves slowly to the positive pole, and the speed of passing through the nanopore is controlled at 1 base per millisecond, and the patch clamp To record the ...

Embodiment 3

[0023] Example 3: Control of perforation speed of cosmid single-stranded DNA

[0024] ① The complex connecting a cosmid single-stranded RNA molecule and a magnetic bead is fixed with an electromagnet at the negative pole of the electrophoresis tank; the two poles of the electrophoresis tank are separated by nanopores.

[0025] ②Turn on the power supply, and give a pulling force (500pN) to the free end of the negatively charged target molecule towards the positive pole. Since the end of the target molecule connected to the magnetic bead is fixed by the electromagnet (the electromagnetic force is 500pN), the result is that the target molecule is straightened. Then through fine-tuning, the electromagnetic force is weakened and the magnetic beads are released, but the electromagnet still attracts the magnetic beads, so that the target molecule moves slowly to the positive pole, and the speed of passing through the nanopore is controlled at 1 base per millisecond, and the patch clam...

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Abstract

The present invention relates to bioengineering technology, and is electromagnetic method and equipment for controlling single-chain nucleic acid perforation. The method includes the following steps: 1. fixing the composition of one single-chain target DNA or RNA and one connected magnetic bead onto the negative pole of electrophoresis tank with two nanometer hole separated poles by using one electromagnet; and 2. turning on the power source to apply the free end of the negatively charged target molecule with one pull force towards the positive pole, trimming the electromagnetic force to release the magnetic bead while maintaining certain attraction of the electromagnet on the magnetic bead so as to make the target molecule to move slowly towards the positive pole and control the nanometer hole passing speed in 0.5-1 base / ms, recording the signal with the patch clamp and converting the signal into sequence information in the computer. The equipment includes electrophoresis tank, nanometer element, computer, etc.

Description

technical field [0001] The present invention relates to a method in the technical field of bioengineering, in particular to a method and device for controlling the perforation speed of single-stranded nucleic acid by electromagnetic force. Background technique [0002] With the advent of the post-genome era, the demand for genome sequencing is increasing dramatically, but the current sequencing methods are slow, expensive, and have gaps. In fact, before the launch of the Human Genome Project, some people began to explore new methods of DNA sequencing. In particular, The National Institutes of Health in the United States launched $1000 Genome in 2004 (that is, in the next 10 years or so, the cost of mammalian sequencing will be reduced. To about $1000, the speed is increased by 3-4 orders of magnitude, the error rate is within 1 / 10,000, and there is basically no gap), and many new sequencing ideas have emerged. Among them, the use of patch clamp to detect single-stranded DNA ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12M1/42G01N27/447
CPCC12Q1/6869C12Q1/6874G01N33/48721
Inventor 王志民
Owner SHANGHAI JIAO TONG UNIV
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