ACTA AGRICULTURAE BOREALI-SINICA ›› 2007, Vol. 22 ›› Issue (6): 24-29. doi: 10.7668/hbnxb.2007.06.006

Special Issue: Biotechnology

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Optimized Method of Preparing and Digesting Megabase-size DNA of Leymus multicaulis Genomes

XU Yue-yu1, ZHOU Yu-lei2, ZHAO Mao-lin3, WANG Zhi-ping4, WANG Ke-wu4, ZHANG Yan1   

  1. 1. Capital Normal University, Beijing 100037, China;
    2. Gansu Agricultural University, Lanzhou 730070, China;

    3. Beijing Academy of Agriculture and Forestry Science, Beijing 100097;
    4. Beijing Soil and Fertilizer Station, Beijing 100029, China
  • Received:2007-03-28 Published:2007-12-28

Abstract: Preparation and digestion of Megabase-size DNA with high quality is the basis for construction of genomic library with large DNA inserts such as yeast articial chromosome(YAC), bacterial artificial chromosome(BAC), binary BAC library(BIBAC), transformation-competent artificial chromosome(TAC), and for long-range physical mapping. This method of embedding the nuclei is optimized to prepare Megabase-size DNA of Leymus multacaulas genomes with high yield and purity. Generally, it includes isolation of purified nuclei by deferential centrifugation, embedment of the nuclei in 1ow-melting-point agarose pulgs, and digestion with proteinase K to release Megabase-size DNA.To purify the high molecular weight DNA with pulsed field gel electrophoresiscan avoid the contaninants of organellar and small molecular weight DNA.Selecting digested DNA fragments‘between 9-300 kb after partial digestion. It proves that the method is suitable for construction of genomic library of Leymus multicaulas.

Key words: Leymus multicaulis, Megabase-size DNA, Genomic library, Pulsed field gel electrophoresis, Digestion

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Cite this article

XU Yue-yu, ZHOU Yu-lei, ZHAO Mao-lin, WANG Zhi-ping, WANG Ke-wu, ZHANG Yan. Optimized Method of Preparing and Digesting Megabase-size DNA of Leymus multicaulis Genomes[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(6): 24-29. doi: 10.7668/hbnxb.2007.06.006.

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