Development and Application of High-Throughput Semi-automatic Genome DNA Extraction Instrument
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1.College of Agronomy, Gansu Agricultural University / Gansu Provincial Key Laboratory of Aridland Crop Science / Gansu Provincial Key Laboratory of Crop Improvement & Germplasm Enhancement;2.Institute of Crop Sciences, Chinese Academy of Agricultural Sciences;3.Biotechnology Research Institute, Chinese Academy of Agricultural Sciences

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Key R&D Program of the Ministry of Science and Technology of China (2019YFD1000700, 2019YFD1000703, 2016YFD0101000, 2016YFD0101001); National Natural Science Foundation of China(31901595)

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    Abstract:

    The high-throughput extraction of high-quality genomic DNA is one of the basic analysis of plant molecular biology research, and an important technology for identifying genomic variation of crop germplasm resources, genetic mapping of important functional genes, crop molecular breeding based on genomic marker-assisted selection. However, the efficiency of conventional method is inadequate for large-scale DNA extraction. The automatic high-throughput instruments are expensive and consumables, and the centrifugal-sedimentation method commonly used in laboratories are tedious and involve more organic solvents. To supply the demand of high-throughput, large-scale and less expensive DNA extraction, we have developed a 96-pool magnetic separation device DNA. Using this device, the genomic DNA extraction has been succeeded for wheat, rice, maize and other cereal crops, as well as vegetable and oil crops. The integrity and purity of DNA were detected by agarose gel electrophoresis, Qubit and PCR. The results showed that (1) the plant genomic DNA had high quality and purity, good integrity; (2) one person can handle 2000 samples per day; (3) the obtained DNA samples can be used in gene cloning, transgenic plant detection, CRISPR editing site detection and NGS library construction.

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History
  • Received:January 23,2022
  • Revised:February 16,2022
  • Adopted:March 09,2022
  • Online: July 08,2022
  • Published:
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