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7种不同种属来源细胞的PCR检测方法与应用

作  者:
韩志玲;郝鹏;商晓桂;范秀丽;闫聪;杨波;刘国英
单  位:
金宇保灵生物药品有限公司
关键词:
细胞;种属;细胞色素b;基因片段
摘  要:
为确定实验室常用细胞来源的可靠性,以及培养过程中是否存在不同种属间细胞的交叉污染现象,本研究针对不同物种的细胞色素b基因进行引物设计并优化,建立PCR扩增体系,对实验室常用的7种不同种属来源的细胞(MDBK、MDCK、BHK21、ST、SP20、293、VERO)进行细胞种属鉴定。试验结果表明:该PCR检测方法特异性较好,操作简便,最低可检测到0.01 ng/μL的DNA含量。结论:本研究所建立的方法能够很好的对7种不同种属的细胞进行鉴别,对细胞培养过程中的质量把控具有重要的指导意义。
译  名:
PCR detection and its application for cells of seven different species
作  者:
HAN Zhiling;HAO Peng;SHANG Xiaogui;FAN Xiuli;YAN Cong;YANG Bo;LIU Guoying;Jinyu Baoling Bio-pharmaceuticals Co.Ltd.;
单  位:
HAN Zhiling%HAO Peng%SHANG Xiaogui%FAN Xiuli%YAN Cong%YANG Bo%LIU Guoying%Jinyu Baoling Bio-pharmaceuticals Co.Ltd.
关键词:
cell;;species;;cytochrome b;;gene fragments
摘  要:
In order to identify the reliability of the cell source and to see whether there is cross-contamination among different cells in the culture process, PCR primers for cytochrome b genes of different species were designed and optimized to perform PCR reactions in seven kinds of cells(MDBK, MDCK, BHK21, ST, SP20, 293, and VERO) in this study. The test results showed that the PCR detection method had good specificity and simple operation, which could detect a minimum of 0.01 ng/μg DNA in content. In conclusion, this method might be used to identify seven different species of cells, providing significant guidance for quality control in the cell culture process.

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