Improved native affinity purification of RNA

RNA affinity tags for purification of RNAs and ribonucleoprotein complexes.

Intrinsic affinity tags are useful tools for the study of macromolecular targets. Although polypeptide affinity tags are routinely used in purification and detection of protein complexes, there has been a relative lack of powerful RNA affinity tags that can be embedded within RNA sequences. Here, the preparation and use of two RNA affinity tags against Sephadex or streptavidin are described. Th...

Sephadex-binding RNA ligands: rapid affinity purification of RNA from complex RNA mixtures.

Sephadex-binding RNA ligands (aptamers) were obtained through in vitro selection. They could be classified into two groups based on their consensus sequences and the aptamers from both groups showed strong binding to Sephadex G-100. One of the highest affinity aptamers, D8, was chosen for further characterization. Aptamer D8 bound to dextran B512, the soluble base material of Sephadex, but not ...

Affinity purification

Stereomicroscope The new Carl Zeiss stereomicroscope Stemi 2000-CS allows the user to view the sample whilst other observers view the procedure by video. Greenough type stereomicroscopes create their stereo effect by having separate light paths set at different angles, one for each eye. However, when video or photomicrography is being carried out most stereomicroscope systems remove light from ...

Improved purification and yields of RNA by RNeasy.

Purification of RNA and RNA-protein complexes by an R17 coat protein affinity method.

We describe an affinity chromatography method to isolate specific RNAs and RNA-protein complexes formed in vivo or in vitro. It exploits the highly selective binding of the coat protein of bacteriophage R17 to a short hairpin in its genomic RNA. RNA containing that hairpin binds to coat protein that has been covalently bound to a solid support. Bound RNA-protein complexes can be eluted with exc...