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Models had SL, temperature, fraction of cells in the S phase, and fraction of cells in the G2/M phases as covariates, and were tested with and without the nRNA covariate included.
Comparison of the RNA fluorescence between frozen tissues and tissues that were frozen and then treated with methanol indicated that short-term methanol preservation did not improve RNA staining; therefore, frozen tissue (stored at -80[degrees]C) worked best for preservation of muscle tissue from larvae of Walleye Pollock for nRNA and DNA staining and was used for all further tests and experiments.
Syto RNASelect stain concentration affected both nRNA and DNA fluorescence.
Therefore, the 1000-nM concentration was optimal for nRNA staining because it produced the highest nRNA fluorescence and had no effect on DNA staining.
The nRNA fluorescence of the RNAse-treated nuclei was significantly less than the values seen for the positive control (DAPI?
Therefore, replicate measurements of nRNA fluorescence from each tank for each treatment were pooled.
Plots of nRNA and DNA fluorescence showed that always-fed larvae had a distinct group of aggregated S-phase nuclei that joined the G0/G1 and G2/M phases, and S-phase nuclei were fewer and dispersed for unfed larvae (Fig.
There was no significant difference in overall nRNA fluorescence between the always-fed and unfed treatments (ANOVA, [F.
There was a loss of DNA when larvae were frozen and thawed, but that process did not affect nRNA measurements.
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- NRMI 2
- NRMI 3