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Process of controlling monochain nucleic acid punching speed by optical nickle

A technology of single-stranded nucleic acid and perforation speed, which is applied in the field of bioengineering and can solve the problem that it cannot be used for actual sequencing.

Inactive Publication Date: 2007-07-18
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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1: Sequencing of single-stranded BAC fragments

[0015] ①The complex of a single-stranded target DNA molecule and a magnetic bead is captured and fixed on the negative pole of the electrophoresis tank with optical tweezers, so that the magnetic beads can withstand the pulling force of 500pN to the negative pole without detaching from the optical tweezers;

[0016] ②Turn on the power supply, and give a pulling force (500pN) from 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 optical tweezers, the result is that the target molecule is straightened, and then moves from the negative pole to the positive pole. Move the optical tweezers to make the target molecules slowly move towards the positive electrode, and control the speed of passing through the nanopore at 0.5-1 base per millisecond. The patch clamp records the signal, and the computer converts the elec...

Embodiment 2

[0018] Example 2: Single-stranded Chromosome Sequencing

[0019] ①The complex of a single-stranded chromosomal DNA molecule and a magnetic bead is captured and fixed on the negative pole of the electrophoresis tank with optical tweezers, so that the magnetic beads can withstand the pulling force of 50pN to the negative pole without detaching from the optical tweezers;

[0020] ②Turn on the power supply, and give a pulling force (50pN) 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 optical tweezers, the result is that the target molecule is straightened, and then moves along the negative pole to the positive pole. Move the optical tweezers to make the target molecules slowly move towards the positive electrode, and control the speed of passing through the nanopore at 0.5-1 base per millisecond. The patch clamp records the signal, and the computer converts the ele...

Embodiment 3

[0022] Example 3: Control of the perforation speed of mRNA

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Abstract

The present invention relates to biotechnology, and is especially optical forceps method of controlling the hole passing rate of single chain nucleic acid. The method includes the following steps: 1. capturing single chain DNA or RNA connected magnetic bead with optical forceps in the negative pole of electrophoresis tank; 2. separating the two poles of the electrophoresis tank with film possessing single nanometer hole so as to perform ion exchange only through the nanometer hole; and 3. turning on the power source for the current to draw the free end of the DNA towards the positive pole, controlling the moving speed of the optical forceps in 0.5-1 base / ms to control the motion of DNA, recording the electric signal during penetrating the nanometer hole with the patch clamp and converting the electric signal into sequence information in the computer. The present invention is superior to available technology, which has too fast nucleic acid speed for patch clamp to recognize.

Description

technical field [0001] The invention relates to a method in the technical field of bioengineering, in particular to a method for controlling the perforation speed of single-stranded nucleic acid by optical tweezers. 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 start 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 the $1000 Genome in 2004 (that is, in the next 10 years or so, the cost of sequencing mammals Since it was reduced to about $1000, the speed was increased by 3-4 orders of magnitude, the error rate was within 1 / 10,000, and there was basically no gap), many new sequencing ideas have emerged. Among them, the use of patch clamp to detect single-stranded DNA (referred to as Sin...

Claims

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Application Information

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