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Preparation and Characterization of a Simulated Cell Free DNA Positive Quality Control Material |
FANG Yan1,2,WANG Xia2,DONG Lian-hua2,YANG Jing-ya1 |
1. College of Food Sciences & Technology, Shanghai Ocean University, Shanghai 201306, China
2. Center for Advanced Measurement Science, National Institute of Metrology, Beijing 100029, China |
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Abstract By optimizing a new type of enzymatic hydrolysis system with unknown mechanism (reaction system, RS), which directly acts on tumor cell lines SW1573,SNU-C2B and NCI-H1975, with KRAS G12C,KRAS G12D,EGFR L858R and EGFR T790M mutations, by optimizing the action time and cell number of RS system, a kind of positive quality control substance which can highly mimic human plasma free DNA (cfDNA) can be prepared by cleavage of cell genomic DNA. The length of simulated cfDNA fragments detected by chip electrophoresis is (166±16)BP, which is the same as that of normal human plasma cfDNA. Microdrop digital PCR detection show that the cfDNA fragments prepared by RS and gDNA have the same mutation abundance at the target mutation site. The results show that the DNA fragment prepared by RS system can be used as a positive quality control substance of cfDNA.
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Received: 17 March 2021
Published: 08 March 2023
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