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These findings demonstrate the superiority of AAV clones selected in vivo over clones selected in vitro, as long as in vivo transduction is required.
The targeting vector was electroporated into C57BL/6 ES cells and clones selected in G418-containing medium.
Then, the infectivity of the best phage clone or clones selected in each of the 0th 20th generations was evaluated simultaneously with the CFU values for JM109.
The mixture was then transformed into DB3.1 and clones selected in the presence of spectinomycin.
This allowed the isolation of 1225 yeast clones, selected in a minimal medium without uracil (Fig. 1).
The RfA-Destination cassette amplicon was transformed into E. coli DB3.1/pWY189/pKD46 induced to express the Red homologous recombination proteins and candidate clones selected in the presence of chloramphenicol and spectinomycin.
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Deletions observed by arrayCGH were confirmed by FISH analysis using one BAC clone selected in each observed deleted region (RP11-734D5 on 11p15.3, RP11-48O9 on 11p13, RP11-545G16 on 11q25).
As the EST analysis progressed, if cDNA clones located upstream of the clones already selected in the contigs appeared, we added these clones into the pipeline for sequencing of the entire inserts.
G418-selected clones were selected in the presence of Epo and then amplified in the presence of Epo or SCF.
Several independent clones were selected in G418 and a clone with low basal and a dose-dependent increase in luciferase activity upon infection with KSHV was selected for further analysis, and designated 293PAN-Luc.
No reduction was found in the clones selected with vincristine.
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