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Antibiotic resistant clones were assayed for Tet-repressor expression by transfecting individual clones with TNF-TetO2-EGFP and monitoring EGFP fluorescence by flow cytometry.
EGY48[p8op-lacZ] was then co-transformed with the different fusion plasmids used in this study and clones were assayed for growth and blue colour onto GR and GRLX media, according to the manufacturer's instructions.
Clones were assayed in the absence and presence of efavirenz and nevirapine (obtained from the NIH AIDS Research and Reference Reagent Program) to monitor RT activity and NNRTI susceptibility.
Surviving single clones were assayed for inducible protein expression of TRAIL.
The clones were assayed at identical protein concentrations and binding detected using anti-FLAG and a tyramine-DNP amplification system.
All five scFv clones were assayed as both monovalent scFv and bivalent scFv-AP at equimolar concentrations (1 × 10-7 M) on HER2 coated wells.
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The growth of the clones was assayed by platting 50,000 cells per well in 12-well plates in triplicates and counted at 1 6 days.
Each clone was assayed in triplicate for all the subsequent follow-up experiments.
Positive clones, isolated using cloning rings, were assayed for SEAP activity using the Great EscAPe SEAP Chemiluminescence Kit 2.0 (Clontech).
Deletion of B19R from the parental NYVAC virus strain did not improve cytokine production by the CD8 T cell clones that were assayed.
Twelve clones from each integration were assayed for self-activation of HIS3 and lacZ expression and the clone with the lowest level for each bait was used for subsequent screens.
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