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- W2373226662 abstract "Objective:To develop a PCR aided transcript titration assay(PATTY) system for quantitation of AFP mRNA . Methods:RT PCR oligonucleotide overlap extension and in vitro transcription were performed with specific AFP primers to construct the mutant fragment of competitor AFP mRNA, AFP mRNAs in hepatoma cell lines were quantitatively analyzed by competitive RT PCR. Results:The mutant fragment of competitor AFP mRNA with a new introduced restriction enzyme site of Hind Ⅲ was constructed and then PATTY system for quantitation of AFP mRNA was developed successfully. AFP mRNA levels in both HepG 2 and Huh7 cell lines were 10 10 copy/μg RNA, in contrast to 10 7 copy/μg RNA in PLC/PRF/5 cell line. Conclusion:The result of quantitating AFP mRNA by this PATTY system is reliable, the competitor AFP mRNA template can supervise the processes of reverse transcription and PCR amplification for detection of target mRNA synchronously. The system may quantitate circulating hepatocellular carcinoma cells associated AFP mRNA . [" @default.
- W2373226662 created "2016-06-24" @default.
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- W2373226662 date "2000-01-01" @default.
- W2373226662 modified "2023-09-25" @default.
- W2373226662 title "A method for quantitation of AFP mRNA by PCR aided transcript titration assay" @default.
- W2373226662 hasPublicationYear "2000" @default.
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