Large-Scale in Vitro Transcription, RNA Purification and Chemical Probing Analysis

Labeling and purification of RNA synthesized by in vitro transcription.

Large scale chemical synthesis, purification and crystallization of RNA-DNA chimeras.

RNA-DNA chimeras, in which both DNA and RNA monomers are site-specifically substituted in the same strand, may be prepared only by chemical synthesis. Biochemical studies have revealed a number of surprising and subtle effects resulting from the insertion of either a ribonucleotide into a DNA strand or a deoxyribonucleotide into an RNA strand. The availability of large quantities of these chime...

Large-scale preparation and purification of polyacrylamide-free RNA oligonucleotides.

We present a fast and simple protocol for large-scale preparation and purification of RNA oligonucleotides. RNA oligonucleotides are prepared by in vitro transcription with T7 RNA polymerase from linearized plasmid DNA templates constructed by PCR. In place of denaturing polyacrylamide gel electrophoresis (PAGE), size-exclusion chromatography is employed to purify the RNA oligonucleotide from t...

Large Scale Identification of SSR Molecular Markers in Ajowan (Trachyspermum ammi) Using RNA Sequencing

The medicinal plant, Trachyspermum ammi is a rich source of active pharmaceutical ingredients with pharmaceutics effects. Microsatellite markers play a key role in the genome and gene expression, especially in secondary metabolite biosynthesis in medicinal plants. For the first time, transcriptome sequencing of this herb medicine was carried out to identify the microsatellite markers of this sp...

Single-molecule correlated chemical probing of RNA.

Complex higher-order RNA structures play critical roles in all facets of gene expression; however, the through-space interaction networks that define tertiary structures and govern sampling of multiple conformations are poorly understood. Here we describe single-molecule RNA structure analysis in which multiple sites of chemical modification are identified in single RNA strands by massively par...