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《Chinese Journal of Blood Transfusion》 2014-06
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Detection of the copy numbers of human coagulation Factor Ⅶ in CHO cell by fluorescent quantitative polymerase chain reaction

XIAO Wei;LI Changqing;LIN Fangzhao;Institute of Blood Transfusion,Chinese Academy of Medical Sciences;  
Objective To detect the copy numbers of Human Coagulation Factor Ⅶ( FⅦ) gene in stable expression CHO cell line with fluorescent quantitative polymerase chain reaction( qPCR). Methods The quantitive genomic DNA of plasmid pMDT-actin and pcDNA-FⅦ were used as templates for making standard curves,respectively. The result was analyzed by Stepone software V2. 2 to calculate the total cell number and the copy number of FⅦ gene in templates. All of the qPCR reactions were performed in ABI Stepone plus system. Results The total cell number in templates is 24 421 ± 7 414.The copy number of the FⅦ gene is 49 455 ± 3 502. No specific gene amplication can be detected in the CHO-pcDNA3. 1.Conclusion The result clearly showed that every CHO cell harbored two FⅦ gene copies.
【CateGory Index】: R440
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