RNA Detection

to living animals. Science 347 (6228):1367–1671. doi:10.1126/science. aaa3380 Haimovich G, Zabezhinsky D, Haas B, Slobo- din B, ...

Chapter 22 Real-Time Fluorescence Imaging of Single-Molecule Endogenous Noncoding RNA in Living Cells Hideaki Yoshimura and Take ...

the difficulty of labeling target RNAs in living cells. RNA cannot be fused with fluorescent or tag proteins through genetic eng ...

PUM-HD mutant; PUM-HD was mutated to address the sequence UUAGGGUU (designated as mPUMt). Telomeres were visualized using a fusi ...

sCMOS camera (e.g., ORCA Flash 4.0 V2; Hamamatsu Pho- tonics K.K.). MetaMorph software (Molecular Devices). 488 nm laser (e.g., ...

acids participating in RNA recognition [8](seeFig. 1d and Note2). In addition to the usage of PUM-HD, employment of a fluo- res ...

to completing the fluorophore formation, takes tens of min- utes, which possibly prevents a precise time-course analysis of the ...

Place the virus-containing mixture to U2OS cells for infection. After 4 h of infection, remove the medium and replace it with f ...

Put all lenses off. Modulate the positions and angles of the lasers and mirrors to make the laser light pass through the center ...

After finding an appropriate cell, fine-tune the focus to an appropriate position for observation of the target RNA. Capture im ...

diaphragm narrow to generate a thin excitation laser beam. When cytoplasmic RNA is the target of the observation, exci- tation l ...

Ozawa T, Yoshimura H, Kim SB (2013) Advances in fluorescence and bioluminescence imaging. Anal Chem 85(2):590–609. doi:10. 1021 ...

Chapter 23 Live Imaging of mRNA Synthesis inDrosophila Hernan G. Garcia and Thomas Gregor Abstract mRNA synthesis is one of the ...

construct design and implementation, embryo handling for live imaging, and various suitable microscopy techniques. We discuss co ...

2 Materials 2.1 Creating MS2 Reporter Constructs and Transgenic Fly Lines Plasmid pCR4-24XMS2SL-stable [4] (Addgene #31865) to ...

2.5 Imaging 1. Custom-built 2-photon microscope [17] with a Zeiss 250.8 NA objective, GaAsP photomultipliers (Hamamatsu H10770P ...

Mix the tube overnight on a nutator. Using forceps extract the tape and pipette the heptane into Eppendorf tubes. Spin down the ...

This sample can be used both on an upright or inverted micro- scope configuration. 3.5 Setting Up the Microscopes for Imaging ...

3.7 Live Data Analysis Download the FlyRNAQuant software and manual. Install the software as instructed in the manual and downl ...

If pronounced photobleaching on the MCP-GFP channel is detected reduce the laser power and/or reduce the frequency of acquisiti ...