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层析凝胶DEAE Sepharose Fast Flow分离纯化5’-混合脱氧单核苷酸
Purification of 5’-deoxynucleotides by DEAE Sepharose Fast Flow
【摘要】 目的针对5’-混合脱氧单核苷酸的理化特性,研究利用DEAE Sepharose Fast F low凝胶分离纯化5’-混合脱氧单核苷酸。方法上样缓冲液采用20 mmol/L Tris,pH9.0,上柱量≤7.5mg/mL.gel,上柱流速5mL/m in,洗脱缓冲液采用20 mmol/LTris+0.025mol/L NaC l,pH 9.0,洗脱流速0.5mL/m in。结果实验结果发现dCMP,dAMP,dTMP的收率为95%以上,dGMP的收率为85%以上,所获得的混合脱氧单核苷酸经高效液相色谱检测色谱纯达98%以上。
【Abstract】 OBJECTIVE According to the physical and chemical characteristic of 5’-deoxymononucleotides,this study investigated the separating and purifying 5’-deoxymononucleotides by DEAE Sepharose Fast Flow.METHODS Take 20mmol/L Tris(pH 9.0) as binding buffer,sample binding≤7.5mg/mL·gel,flow-rate 5mL/min;Take 20mmol/L,0.025mol/L NaCl(pH 9.0) as elution buffer,flow-rate 0.5mL/min. RESULTS The chromatographic grade purity of the eluate is above 98% by HPLC,the recovery rate of dCMP,dAMP and dTMP is above 95%,the recovery rate of dGMP is above 85%.
【Key words】 5’-deoxymononucleotides; DEAE Sepharose Fast Flow; recovery rate; purity;
- 【文献出处】 中国现代应用药学 ,Chinese Journal of Modern Applied Pharmacy , 编辑部邮箱 ,2007年01期
- 【分类号】TQ464.6
- 【被引频次】4
- 【下载频次】476