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Extraction method of genome DNA of isolated soy proteins

A soybean protein isolate and extraction method technology, applied in the field of genomic DNA extraction, can solve problems such as inability to extract genomic DNA, and achieve the effects of facilitating rapid detection, simple operation, and short time-consuming

Inactive Publication Date: 2011-10-05
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to solve the existing soybean genome DNA extraction method with soybean seedling leaves, soybean dry seeds, and fresh pods at the bulging stage as materials. The above-mentioned extraction method cannot be extracted from protein-rich soybean protein isolates. The problem of genomic DNA, and then provide a method for extracting genomic DNA of soybean protein isolate

Method used

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Embodiment Construction

[0006] The preferred embodiment of the present invention is as follows: firstly, the soybean protein isolate and TE buffer solution are fully mixed to form a TE mixed solution, and then the guanidine isothiocyanate lysate with twice the volume of the mixed solution is added, fully mixed, and lysed at room temperature. Add an equal volume of phenol, chloroform / isoamyl alcohol to extract protein, add an equal volume of chloroform / isoamyl alcohol to the supernatant after centrifugation to extract protein, add an equal volume of chloroform to the supernatant to extract protein after centrifugation, and centrifuge , the supernatant was precipitated with isopropanol, centrifuged, the precipitate was washed with ethanol, dried at room temperature, and dissolved in TE buffer to obtain a TE solution of genomic DNA.

[0007] The soybean protein isolate in the TE mixed solution is 30-80 mg, and the TE buffer solution is 120-320 μL. The preferred range is 50 mg of soybean protein isolate ...

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Abstract

The invention discloses an extraction method of the genome DNA of isolated soy proteins, comprising the following steps of: firstly sufficiently mixing isolated soy proteins and a TE buffer solution to prepare a TE mixed solution; then adding a guanidinium isothiocyanate lysis solution with volume twice larger than that of the TE mixed solution, sufficiently uniformly mixing, and lysing at room temperature; adding isometric phenol and chloroform / isoamylol to extract proteins; centrifugalizing, and then adding isometric chloroform / isoamylol to a supernate to extract the proteins; centrifugalizing, and then adding isometric chloroform to the supernate to extract the proteins; centrifugalizing, and precipitating the supernate by using isopropanol; centrifugalizing, and washing by using ethanol for precipitation; airing at the room temperature; and dissolving in the TE buffer solution so as to obtain the TE solution of the genome DNA. According to the extraction method, the genome DNA which has high quality and is suitable for PCR (Polymerase Chain Reaction) detection is extracted from the isolated soy proteins, the concentration of the genome DNA is 1558 micrograms / ml, and the value of OD260 / OD280 is 1.666; and in addition, the invention has the advantages of easy operation and short consuming time and is beneficial to fast detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for extracting genome DNA of isolated soybean protein which is suitable for PCR amplification. Background technique [0002] At present, there have been many studies and reports on the extraction of plant genome DNA, but due to the different components of different plants or different tissue materials of the same plant, the appropriate extraction methods are also different. The high protein content of soybean protein isolate is not conducive to the dissolution and extraction of DNA. General plant genomic DNA extraction methods cannot extract genomic DNA from soybean protein isolate, which brings difficulties for the next step of gene-based biological detection. The existing soybean genomic DNA extraction method uses soybean seedling leaves, soybean dry seeds, and fresh pods at the bulge stage as materials for extraction. The above extraction methods cannot extract genomic DNA...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 徐伟丽马莺李启明
Owner HARBIN INST OF TECH
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