RNA Detection

(GUI). Alternatively, edit the calibration settings in the Track- Mate GUI. Make sure that both image series are calibrated the ...

Examine all tracks to see whether they describe spot movement accurately. If not, go back and change tracking parameters accord ...

3.4.2 Track Colocalization Open the Track-based colocalization analysis workflow in KNIME. Configure and execute the “Parameter ...

Inspect the interactive table generated by “JavaScript Table View” node (#426). It contains the results of the colo- calization ...

3.4.4 Output 1. Inspect the results of the translation analysis that are saved in a “KNIME_Results.xlsx” file in the parent fold ...

Acknowledgments The authors would like to thank all members of the Chao lab for their joint efforts in developing the TRICK anal ...

Chapter 26 Imaging Translation Dynamics of Single mRNA Molecules in Live Cells Suzan Ruijtenberg, Tim A. Hoek, Xiaowei Yan, and ...

Here, we describe a microscopy-based method, which allows quantitative measurements of ribosome initiation and elongation on ind ...

Fig. 1Imaging translation of single mRNA molecules. (a) Schematic representation of the fluorescent labeling of nascent polypept ...

Cell type specific cell culture medium. Live-cell imaging cell culture medium; we use Leibovitz’s-L15 imaging medium (Gibco, Li ...

Alternatively, start with a vector of choice and a POI of choice and clone the individual elements of the translational reporte ...

3.1.2 sfGFP-Tagged Antibody that Binds the SunTag Peptide (scFv-GFP) The scFv-GFP binds with high affinity to the SunTag peptide ...

binding sites and the sequence encoding the SunTag peptides), virus titers may be low, and transfection may be preferred over vi ...

keeping a lid on the imaging plate whenever possible to prevent evaporation. When using a doxycycline-inducible promoter to exp ...

Selectthe appropriatemicroscope objective. In general, we image with a 100
NA1.49 oil-immersion objective to obtain a high reso ...

3.5 Drugs that Interfere with Translation as Tools to Study Translation Dynamics Several drugs that are known to interfere with ...

concentration of harringtonine in DMSO, make a dilution of 21 μg/mL (7
the final concentration) in imaging medium. Of this dilut ...

(d) Subtract the mean background intensity from the mean spot intensity to obtain the intensity of a single SunTag protein. Mea ...

that other regulatory sequences outside the coding region, such as the 3^0 and 5^0 UTR, may influence translation efficiency as ...

is, however, an important parameter to take into account. The longer the reporter gene, the more ribosomes can be present on the ...