Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (6): 74-80.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1123

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Rapid Crude Extraction of Genomic DNA from Solanum lycopersicum for PCR

SHEN Heng1(), LIU Si-hui2, LI yue1, LI Jing-tao1, LIANG Wen-xing1()   

  1. 1. Key Lab of Integrated Crop Pest Management of Shandong Province,College of Plant Health and Medicine,Qingdao Agricultural University,Qingdao 266109
    2. College of Science and Information,Qingdao Agricultural University,Qingdao 266109
  • Received:2021-08-31 Online:2022-06-26 Published:2022-07-11
  • Contact: LIANG Wen-xing E-mail:2963461817@qq.com;wliang1@qau.edu.cn

Abstract:

It is necessary to extract plant genomic DNA and perform PCR cloning when conducting the studies of molecular biology on plants. Currently,the CTAB(Cetyltrimethyl Ammonium Bromide)is commonly used to extract plant DNA in laboratories,but the steps are relatively cumbersome and require multiple organic solvents. Specifically,it takes a long time when there are many samples. The DNAs from multi-species were extracted by NaOH,and this study introduces a method of extracting plant genome DNA,which is faster,economical,safer and more efficient. The major steps include the followings:Quickly grinding the plant tissues using liquid nitrogen;quickly extracting DNA by alkaline lysis(0.5 mol/L NaOH);neutralizing the extracted DNA sample with TE buffer;and performing PCR with neutralized DNA as template. Using this simple DNA extraction method and combining PCR detection,the rapid preliminary identification of transgenic S. lycopersicum was completed. Furthermore,it was confirmed that this NaOH method could be used for extracting DNA from the different tissues of S. lycopersicum. The entire extraction process was simple and of short-time. Thereby,DNA extraction for a large number of plant samples could be completed in a short time without using toxic regents such as chloroform and isopropanol etc. The completion time of DNA extraction is about 1/5 of the time while using CTAB method. In addition,the extracted DNA from different tissues are of good integrity and high quality,which meet regular PCR detection,which can be used for gene cloning and for transgenic tomato screening. This method demonstrates potential utilization value and prospects.

Key words: Solanum lycopersicum, NaOH, DNA extraction, PCR, identification of transformant