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

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。

OBJECTIVE According to the physical and chemical characteristic of 5＇-deoxymononudeotides, 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%.