RNA Detection

peptides emerging from the ribosome are labeled incompletely, limiting the fluorescence of translation sites. Tethering the mRN ...

Transient versus stable transfection. In general, the plasmids required for translation imaging (seeSubheading2.1 )canbe either ...

which represent sites of translation (Fig.3). scFv-GFP expres- sion levels need to be sufficiently high to bind all SunTag pep- ...

Advantages and disadvantages of different imaging systems. Widefield microscopy: Widefield microscopy allows imaging of relativ ...

will thus localize slightly further toward the cell interior (Fig.1a, lower panel). Therefore, care should be taken to ensure bo ...

into account. A simple mathematical model can be used to calculate the number of ribosomes from the measured GFP intensities [5] ...

Chapter 27 Systematic Detection of Poly(A) + RNA-Interacting Proteins and Their Differential Binding Miha Milek and Markus Landt ...

Compared to other gene expression regulators such as tran- scription factors, RNA-binding proteins (RBPs) show higher expression ...

Rotating wheel for falcon tubes. Liquid nitrogen. 2.2 Solutions We routinely use distilled water (MilliQ grade) to prepare the ...

Nonlabeled/“light” cell culture medium: mix 0.5 L of Dulbecco’s modified Eagle’s medium (DMEM) containing high glucose (4.5 g/L ...

0.4 M Arg10 solution: Reconstitute 62.1 mg of^13 C 6 ,^15 N 4 L- arginine HCl (Cambridge Isotope Laboratories) in 700μL deioniz ...

Pass the lysate with a syringe several times through a 23- and 26-G needle (seeNote 4). Afterward, save 0.2 mL of input lysate ...

Keep the eluates on ice or store them at 80 C until further use. Wash and store oligo(dT) beads in lysis/binding buffer at 4 ...

ice-cold PBS. Collect cells and freeze pellets in liquid nitrogen or continue with lysis (seeNote 19). Lyse ‘light” and “heavy” ...

Analyze the oligo(dT) eluates as described in Subheading3.3. Representative results are shown in Fig.2b. Submit the sample for ...

separating it from the top viscous layer. To avoid RNA frag- mentation, sonication is not recommended. We suggest to use 200μL ...

volume of water. Note that due to denaturation some proteins cannot be effectively resuspended after TCA precipitation. After r ...

The amount of spiked-in volume should be discussed with the scientist performing the mass spectrometry experiments. After recei ...

Liepelt A, Naarmann-de Vries IS, Simons N, Eichelbaum K, Fohr S, Archer SK, Castello A, Usadel B, Krijgsveld J, Preiss T, Marx ...

Chapter 28 Isolation and Characterization of Endogenous RNPs from Brain Tissues Rico Schieweck, Foong yee Ang, Renate Fritzsche, ...