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Detection of genomically integrated RNAi-cassettes in HeLa cell clones
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posted on 2011-12-30, 16:42 authored by Sven Kappel, Yves Matthess, Brigitte Zimmer, Manfred Kaufmann, Klaus StrebhardtCopyright information:
Taken from "Tumor inhibition by genomically integrated inducible RNAi-cassettes"
Nucleic Acids Research 2006;34(16):4527-4536.
Published online 31 Aug 2006
PMCID:PMC1636372.
© 2006 The Author(s)
Genomic DNA from HeLa cell clones digested with HindIII was further analyzed. () PCR was performed to amplify a sequence of 185 bp in length enclosing different H1 promoter derivatives and shRNA-coding regions. Subsequent sequencing of amplified fragments revealed the authentical integration of different RNAi-cassettes. DNA from wild-type HeLa cells was used as negative control (lane 7), recombinant plasmid DNA was used as positive control (lane 6). () Southern blot analysis was performed to determine the number of integrated RNAi-cassettes (arrows). A radioactive probe specific for RNAi-cassettes was used.