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This measurement captures sampling variability as a result of particle aggregation, allowing for normalization based on iron concentration in each aliquot of aggregate suspension added to the experimental setup.
Cells were also co-transfected with a CAT expression construct, allowing for normalization.
Unlabeled reporter peptide R9L was included in each assay as competitor to allow for normalization of data between different assays.
In both cases, subtracted cDNAs were subjected to two PCR cycles to allow for normalization of cDNA populations.
PCR amplification of 18S RNA was done for each sample as a control for sample loading and to allow for normalization between samples.
Additionally, expression of the 18S ribosomal subunit was also assessed to allow for normalization of total RNA levels between samples using the 2−ΔΔCT method [38].
The gels were subsequently stained with Deep Purple total protein stain to allow for normalization of the Pro-Q Diamond signal according to total MYBP-C content.
Even though 60-fold more vge/cell was utilized with HPV18 compared to HPV45, this allowed for normalization of RT-qPCR data, allowing amplification of nucleic acid at similar Ct values (data not shown).
Even though 10-fold more vge/cell was used with HPV31 compared to HPV16, this allowed for normalization of RT-qPCR data, allowing amplification of nucleic acid at similar Ct values (data not shown).
We used cell-line controls to span the dynamic range of the measurements, allowing for normalization of the data across multiple batches and ensuring our ability to compare different batches.
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