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Kit for extracting bacterial plasmid DNA (deoxyribonucleic acid) by magnetic beads, and extraction method thereof

A bacterial plasmid and extraction method technology, applied in the field of molecular biology, can solve the problems of high-speed centrifugation, vacuum filtration column separation and automatic extraction, and no extraction of bacterial plasmid DNA, etc., to achieve low price, short extraction time and high purity Effect

Inactive Publication Date: 2011-11-16
HENAN HUIER NANO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the existing commercialized plasmid DNA extraction kits have the advantage of no need for organic solvent phenol-form extraction, which is much better than traditional extraction methods, but they still cannot get rid of the tedious high-speed centrifugation, vacuum filtration, column separation and inability to use Defects in automatic extraction of instruments
However, there is no relevant technology and report about the magnetic bead method to extract bacterial plasmid DNA.

Method used

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  • Kit for extracting bacterial plasmid DNA (deoxyribonucleic acid) by magnetic beads, and extraction method thereof
  • Kit for extracting bacterial plasmid DNA (deoxyribonucleic acid) by magnetic beads, and extraction method thereof

Examples

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Effect test

Embodiment 1

[0034] A kit extraction method for extracting bacterial plasmid DNA by a magnetic bead method, which specifically comprises the following steps:

[0035] Step 1. Take 1.0ml of bacterial culture solution cultivated overnight, add it to a 1.5ml centrifuge tube (EP), centrifuge at 12000 r / min for 7 minutes, discard the supernatant, keep the bacteria, add 50μl of resuspension solution, and vortex for 5 Seconds, resuspend the bacteria to ensure that there are no visible small pieces after resuspension;

[0036] Step 2. Add 100 μl of lysate to the resuspended cells, invert the centrifuge tube 5 times to mix well, and continue for 2 minutes. Do not vibrate vigorously, otherwise the genomic DNA will be sheared until the solution becomes viscous but clear;

[0037] Step 3. Add 75 μl of neutralizing solution to the lysate obtained in step 2, and immediately invert the centrifuge tube 5 times until flocculent precipitates appear in the solution. Centrifuge at 12,000 r / min for 10 minutes ...

Embodiment 2

[0042] A kit extraction method for extracting bacterial plasmid DNA by a magnetic bead method, which specifically comprises the following steps:

[0043] Step 1. Take 1.5ml of bacterial culture solution cultivated overnight, add it to a 2.0ml centrifuge tube (EP), centrifuge at 13000 r / min for 10 minutes, discard the supernatant, keep the bacteria, add 65μl of resuspension, and vortex for 6 Seconds, resuspend the bacteria to ensure that there are no visible small pieces after resuspension;

[0044]Step 2. Add 130 μl of lysate to the resuspended cells, invert the centrifuge tube 5 times to mix, and continue for 3 minutes. Do not vibrate vigorously, otherwise the genomic DNA will be sheared until the solution becomes viscous but clear;

[0045] Step 3. Add 100 μl of neutralizing solution to the lysate obtained in step 2, and immediately invert the centrifuge tube 5 times until flocculent precipitates appear in the solution. Centrifuge at 13,000 r / min for 10 minutes to form a com...

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Abstract

The invention discloses a kit for extracting bacterial plasmid DNA (deoxyribonucleic acid) by magnetic beads, and an extraction method thereof. The kit comprises six components: a bacterial resuspension solution, a cracking solution, a neutralizing solution, a magnetic bead combining solution, a washing solution and an eluting solution. The extraction method of the kit comprises the following steps of: resuspending bacteria, cracking, neutralizing, combining with magnetic beads, washing and eluting. The plasmid DNA extracted by the invention has high purity and complete fragments, and the extraction process is convenient, quick, efficient and safe.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a kit for extracting bacterial plasmid DNA by a magnetic bead method and an extraction method thereof. Background technique [0002] The birth and development of molecular biology is a major event in the entire natural sciences, which brings the research of life sciences to a new stage. DNA is not only the basic genetic material, but also plays a decisive role in the synthesis of proteins and the growth and variation of a series of major life. Whether it is genetic engineering or protein engineering, the DNA must first be extracted and purified, especially in genetic engineering, the extraction of bacterial plasmid DNA is very important. [0003] At present, the methods commonly used to extract plasmid DNA from bacteria include alkali lysis, boiling, SDS lysis, etc. These methods mainly consist of bacterial culture (amplification of plasmid DNA), bacterial lysis (relea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 王平康杜德光宋昕孙荣青李祺曹露聪
Owner HENAN HUIER NANO TECH
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