Acta Chimica Sinica ›› 2001, Vol. 59 ›› Issue (9): 1464-1470. Previous Articles     Next Articles

Original Articles

HPLC技术对细菌基因组DNA碱基组成的精确测定

李越中;胡玮;张禹清;张勇;韩冠君;黎志凤;王冰   

  1. 山东大学生命科学系微生物技术国家重点实验室
  • 发布日期:2001-09-15

Precise determination of base composition in bacterial genome DNA using HPLC techniques

Li Yuezhong;Hu Wei;Zhang Yuqing;Zhang Yong;Han Guanjun;Li Zhifeng;Wang Bing   

  • Published:2001-09-15

Errors of reproducibility with high performance liquid chromatography (HPLC) are remarkably serious in determination of base composition of bacterial genome DNA, which is usually defined as guanine-plus-cytosine content [(G+C) mol%]. For partially purified DNA, salts are a major source of interference in enzymatic degradation and RNAs are a major source of interference in chemical degradation. This research analyzes the conditions and products by chemical (perchloric acid and formic acid) and enzymatic degradations (phasphodiesterase Ⅰ and bovine intestinal mucosa alkaline phophatase) of bacterial genome DNA using RP-HPLC, and introduces a systematic method to precisely measure the base composition of bacterial DNA.

Key words: HIGH SPEED LIQUID CHROMATOGRAPHY, HYDROLYSIS, PERCHLORIC ACID, FORMIC ACID, PHOSPHODIESTERASE, PHOSPHOKINASE

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