To determine the rate-limiting step, we used nucleotide labelled with the fluorescent moiety mant (methylanthraniloyl) (Supplementary Figure S2A).
The assay is essentially the same as the one described by Kim et al. [ 6], but, instead of radioactive nucleotide labeling, biotinylated primers were used during the extension reaction, thus enabling quantitative photometric detection by a streptavidin-DIG-peroxidase immunoreaction.
The IVT reaction for amplification and labeling of the target is similar in most aspects, except the BioArray method uses two nucleotide labels (biotin-CTP and biotin-UTP), the Superscript kit a single biotin-UTP labeling and the One-Cycle, a ψ-UTP biotinylated nucleotide (Table 3).
These include the expression of cell surface or intracellular markers, mammosphere formation, exclusion of fluorescent dye by a SP and the retention of the radio-nucleotide label.
These mutant proteins were analysed for their interaction with guanine nucleotides labeled with a fluorescence dye and for their ability to hydrolyse GTP in a cooperative manner.
The method includes the use of modified nucleotides labeled radioactively or by fluorescence and gel electrophoresis [95].
In fact, the percentage of nucleotides labeled as a COUP-TF HMM match was fairly constant throughout the genome (Figure 2D).
Conserved nucleotides (labelled "1" and "4" in Figure 5B) in these −10 and −35 motifs were then mutated and the mutated fragments were individually re-cloned into pMP220.
Then, four types of nucleotides labeled by four colors are added in, and sequencing is performed via the sequencing by synthesis (SBS) method.
Next, the T7 polymerase was added along with nucleotides labeled with fluorescent cyanine-3 (Cy3) dye, which amplified the anti-sense complementary RNAs (cRNA).
