Method for simple and rapid high-throughout extraction of plant genome DNA

An extraction method and whole genome technology, applied in the field of DNA extraction, can solve the problems of cumbersome and expensive, unfavorable population genetics, high cost, etc., and achieve the effect of low cost

Inactive Publication Date: 2012-12-26
匡贤彦 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Currently (2011) Qiagen Puregene Accessories on the market include Cell Lysis Solution (1000ml), Protein Precipitation Solution (350ml) and DNA Hydration Solution (500ml) kit quality and effect are not stable due to different production batches, and the purification process is relatively complex Long (5 hours), high cost (3,000 US dollars, about 1 US dollar / DNA sample), and its cumbersome and expensive defects are very unfavorable for population genetics, phylogenetic evolution, molecular marker-assisted breeding, and deep sequencing.

Method used

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  • Method for simple and rapid high-throughout extraction of plant genome DNA
  • Method for simple and rapid high-throughout extraction of plant genome DNA
  • Method for simple and rapid high-throughout extraction of plant genome DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046]Embodiment one: use a puncher to punch a circular blade with a diameter of 0.6 cm on the fresh corn blade. Leaves can be stored at 4 degrees Celsius for later use, extracted within 1 week, or dried leaves stored at room temperature for a long time. Put the leaves, 1 steel ball with a diameter of 0.4 cm and 200 microliters of extraction solution in a small tube of a 96-well plate. Place a layer of Parafilm sealing film on the 96-well plate, then press the cover of the 96-well plate tightly, and press the tops of the 96 small tubes one by one with a flat surface (about 1 cm in diameter) of a hard object to prevent the extraction solution and tissue fluid from seeping. leak. Place the 96-well plate on the shaker, set the frequency to 30 times / second, shake for 30 seconds, rotate the 96-well plate 180 degrees, put it on the shaker again, and shake at the same frequency for another 30 seconds. Put the shaken 96-well plate into a centrifuge, set the speed at 5000 rpm, and ce...

Embodiment 2

[0047] Embodiment two: use a puncher to punch a circular blade with a diameter of 0.6 cm on the fresh rice blade. Leaves can be stored at 4 degrees Celsius for later use, extracted within 1 week, or dried leaves stored at room temperature for a long time. Other steps are the same as in Embodiment 1. QiagenPurgene was used to extract DNA as a control.

Embodiment 3

[0048] Embodiment three: use a puncher to punch a circular blade with a diameter of 0.6 cm on the fresh sorghum blade. Leaves can be stored at 4 degrees Celsius for later use, extracted within 1 week, or dried leaves stored at room temperature for a long time. Other steps are the same as in Embodiment 1. Use Qiagen

[0049] Purgene DNA extraction method was used as a control.

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Abstract

The present invention provides a method for simple and rapid high-throughput extraction of plant genome DNA, and relates to the technical field of a plant complete gene DNA extraction method. A purpose of the present invention is to solve technical problems of high cost, complex operation, low throughput, long time consuming, low purity, easy pollution, and the like in the traditional and existing (2011) plant genome DNA extraction method. According to the present invention, self-preparing reagent formulas are adopted, and an enzyme reagent is not adopted, such that cost is low; liquid nitrogen grinding is not required, such that operation is simple; 384 samples can be performed in one time, such that throughout is high; an extraction process requires only 2.5 hours, such that a consumed time is short; only 0.5 mul of the extracted DNA is required to be adopted as a PCR template, such that purity is high; and a Parafilm film is adopted for sealing, such that characteristics of sealing, leakage resistance, cleaning, and no pollution are provided. In addition, the extracted plant complete genome DNA can be applicable for plant population genetics, phyletic evolution, molecular marker-assisted breeding, deep sequencing, and other researches, and suitable for industrial production and ordinary biological laboratory science researches.

Description

technical field [0001] The invention relates to a DNA extraction method, in particular to a plant whole gene DNA extraction method. Background technique [0002] Plant cells not only have high nuclease activity, but also contain a large number of polysaccharides in their cell walls, and their composition varies with species. These polysaccharides affect the co-precipitation step in the extraction process of plant genomic DNA and reduce the purity of nucleic acid. The routinely used CTAB method, urea extraction method, and SDS / phenol / chloroform extraction method for genomic DNA are not only complex and time-consuming, but also use a large amount of volatile organic compounds and generate a large amount of toxic waste liquid. [0003] Currently (2011) Qiagen Puregene Accessories on the market include Cell Lysis Solution (1000ml), Protein Precipitation Solution (350ml) and DNA Hydration Solution (500ml) kit quality and effect are not stable due to different production batches,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 不公告发明人
Owner 匡贤彦
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