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Putative phosphopeptides (neutral loss of H3PO4) reported by Mascot searches ("Phosphorylation (ST)" was used as variable modification) were verified by manual inspection of the corresponding MS2 spectra.
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Performance of the modification was verified taking various experiments.
While increase in cellular interaction was clearly observed from the CCK-8 and histological stain data, its surface modification was verified by various physical and chemical changes.
The proposed modification was verified against the test results of an extensive experimental work conducted at the University of Sherbrooke on full-scale two-way specimens measuring 2500 mm × 2500 mm × 200 mm or 350 mm.
LDL modification was verified by particle migration on the Paragon® Electrophoresis System (Beckman Coulter, Fullerton, CA).
Briefly, lipoproteins (5 µg protein each) were loaded onto the gel and subjected to electrophoresis for 30 min at 100 V. Particle modification was verified by migration of the oxLDL relative to native LDL (N-LDL) samples.
DNA locus modification was verified by PCR with primers P9 – P90.
After Red recombination, the DNA locus modification was verified by PCR, and followed by λ-Int/Xis-mediated excision of the marker from the chromosome.
After two rounds of homologous recombination, engineered C. glutamicum with the corresponding chromosomal modifications were verified by PCR.
The modifications were verified by DNA sequencing (in-house ABI lab DNA sequencing core facility, Dept. Molecular Biosciences, University of Oslo).
All constructs were generated by restriction and modification enzymes and all cloning modifications were verified by sequencing.
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