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Method of separating plastid DNA employing silicon oxide-magnetite nano composite

A nanocomposite, magnetite technology, applied in the field of separating plastid DNA

Inactive Publication Date: 2007-02-07
邱良瑛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, to date, silica has not been utilized to develop similar preparations for good-quality plastid DNA.

Method used

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Experimental program
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Effect test

Embodiment 1

[0044] Material:

[0045] Ferric chloride hexahydrate was purchased from J.T. Baker (Phillipsburg, NJ, USA). Ferrous chloride tetrahydrate was purchased from Fluka (Buchs, Switzerland). PEI, tetraethoxysilane, and lauric acid were all supplied by Sigma Chemical Co. (St. Louis, MO, USA). Sepharose L (low osmolarity) was obtained from Amersham Biosciences (Uppsala, Sweden). The supercoiled DNA ladder was obtained from Invitrogen Co. (Carlsbad, CA, USA). The reagents used in DNA isolation and analysis are of molecular biology grade. RNase A was obtained from Sigma. All other chemicals and solvents used were of analytical grade and used without further purification. The water used in the whole experiment was produced by Milli-Q Ultra-Pure-Water Purification System of Nihon Millipore Ltd. (Tokyo, Japan). All solutions were freshly prepared.

[0046] method:

[0047] 1. Plastid composition

[0048] The complete enhanced green fluorescent protein gene (EGFP) was amplified fr...

Embodiment 2

[0073] 1. Identification of magnetic particles

[0074] The size and morphology of the magnetic nanoparticles were identified by TEM. figure 1 Typical TEM micrograph showing magnetic particles. The superparamagnetic properties of the magnetic particles are confirmed by the magnetization curve measured by SQUID. Figure 2 shows a typical plot (M-H loop) of magnetization versus applied magnetic field at 298K. The resulting Fe 3 o 4 The saturation magnetization of the magnetic particles is 64emu / g Fe 3 o 4 . The large saturation magnetization of these magnetic particles makes them very sensitive to magnetic fields, thus allowing easy separation of solid and liquid phases. Very weak hysteresis reveals that the resulting magnetic nanoparticles are nearly superparamagnetic.

[0075] 2. The influence of ionic strength

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Abstract

the invention discloses a separating method of plastid DNA through silica-magnetic nanometer compound, which comprises the following steps: preparing superparamagnetism ferric oxide nanometer particle based on Fe2+ and Fe3+ salt and ammonium hydroxide under nitrogen environment through chemical sedimenting method; hydrolyzing tetraethoxysilane through acid to produce silica on the magnetic ferric oxide nanometer particle to prepare silica-magnetic nanometer compound carrier; adsorbing sample with plastid DNA and silica-magnetic nanometer compound carrier on the carrier under high-salt condition; recycling the silica-magnetic nanometer compound carrier through permanent magnet; recycling the plastid DNA through remained clarified solution.

Description

1. Technical field [0001] The invention relates to a method for isolating plastid DNA, more particularly to a method for isolating ultrapure plastid DNA from bacterial culture by using a silicon oxide-magnetite nanocomposite. 2. Background technology [0002] Purification of specific cells or molecules is fundamental to life science research, but some isolation methods can be physically and chemically hazardous. In the current state of the art, magnetic separation offers a milder alternative, where the target is captured on magnetic particles coated with a target-specific surface and a magnetic field is used to separate the target from the sample. For rapid isolation of bacterial plastid DNA, the need for methods to free the isolated DNA from proteins, RNA, salts, and enzyme inhibitors has increased dramatically with the proliferation of molecular experimental procedures requiring highly purified gene templates. The most prevalent plastid isolation methods are quite time-co...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 江祯立宋锦珊
Owner 邱良瑛
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