RNA Detection

Chen H, Xu Z, Mei C, Yu D, Small S (2012) A system of repressor gradients spatially orga- nizes the boundaries of bicoid-depend ...

Chapter 24 Imaging Newly Transcribed RNA in Cells by Using a Clickable Azide-Modified UTP Analog Anupam A. Sawant, Sanjeev Galan ...

CuAAC, strain-promoted azide–alkyne cycloaddition (SPAAC) and azide–phosphine Staudinger ligation reactions. Although the ver- s ...

autoclaved water. Sterilize by filtering HBS solution using sterile 0.2μm syringe filter inside the cell culture hood and store ...

50 mL Falcon using sterile serological pipette. Add 5 mL of heat inactivated FBS using sterile serological pipette and gently mi ...

2.3 Reagents for Azide–Alkyne Cycloaddition Reaction 7.5 mM Alexa594 alkyne: Prepare a stock solution of Alexa Fluor®594 alkyne ...

3.1 Transfection of AMUTP into Cells Culture mammalian cells (e.g., HeLa cells) in complete DMEM supplemented with 10% FBS (see ...

3.2 Imaging RNA in Fixed Cells by Using CuAAC Reaction After transfecting AMUTP for 1 h, remove the transfection mix. Wash the ...

3.4 Imaging RNA in Live Cells by Using SPAAC Reaction Culture HeLa cells in complete DMEM supplemented with 10% FBS (seeNote 1) ...

Gently swirl the dish in a clockwise and anticlockwise direction so that the transfection mix covers the surface of the cells. ...

After SPAAC staining (30 min at 37C), remove the media and add 1 mL of Hoechst 33342 (1μM) prepared in complete DMEM to the ce ...

4 Notes Thaw buffer solutions and media required for cell culture in the water bath (37C) containing autoclave water prior to ...

avoid extended exposure to light. Dye stocks can be either wrapped using aluminum foil or stored in amber-colored tubes. Degass ...

References El-Sagheer AH, Brown T (2010) New strategy for the synthesis of chemically modified RNA constructs exemplified by ha ...

Chapter 25 Detection of the First Round of Translation: The TRICK Assay Franka Voigt, Jan Eglinger, and Jeffrey A. Chao Abstract ...

associated with the coding sequence of an mRNA transcript during the first round of translation (Fig.1a). A TRICK reporter trans ...

loops can be bound by PP7 and MS2 coat proteins (PCP and MCP) fused to spectrally distinct fluorescent proteins, an untranslated ...

a dichroic beam-splitter in the scanhead (Semrock Di01- T488/568—13  15  0.5), 100 1.45NA PlanApo TIRFM oil immersion objecti ...

(BamHI/ClaI) cassettes can be individually cloned into reporter constructs. Conventional PCR amplification of the PP7 and MS2 ca ...

which would prevent accurate spot detection in the last frames of the image series. Camera gain can be adjusted with respect to ...