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To confirm Stx phage transfer by bAGPs, we randomly chose transductant colonies from the co-culture experiment described earlier (Figure 3) for further verification with PCR to detect genes specific for the Stx2 phage (stx2), E.
We calculated the transduction frequencies by dividing the number of transductants by the number of recipients.
Pink colonies growing on sorbitol-MacConkey agar supplemented with either kanamycin and tetracycline or kanamycin and ampicillin were regarded as presumptive transductants (i.
Generation of OSC19 Transductant Lines and Cell Culture Conditions
The generation of stable UMSCC1 control and precursor miR375-expressing (Pre375) lentiviral transductant cell lines was previously described.
UMSCC1 transductant cells were lysed in radioimmunoprecipitation assay buffer containing 1X protease inhibitors (Cat.
UMSCC1 transductant cells were lysed with cold IP Lysis/Wash Buffer containing phosphatase inhibitor cocktails (set 1: Cat.
The numbers of primary clones and transductant clones amplified were 2 x [10.