Gene expression profiling from microdissected cell populations is a powerful approach to explore molecular processes involved in development and solid tumor biology. In this chapter, we detail robust and validated methods for tissue preparation and isolation of high-quality RNA from microdissected cell populations. A protocol is also provided for linear transcript amplification using as little ...

The impact of three different RNA isolation methods on the community analysis of metabolically active bacteria was determined by reverse transcription (RT) and PCR amplification of 16S rRNA genes and subsequent terminal restriction fragment length polymorphism (T-RFLP) analysis. Furthermore, soil samples were stored at different conditions in order to evaluate the effect of soil conservation me...

Pseudomonas aeruginosa (PA) infections result in significant morbidity and mortality in hosts with compromised immune systems, such as patients with leukemia, severe burn wounds, or organ transplants. In patients at high-risk for developing PA bloodstream infections, the gastrointestinal (GI) tract is the main reservoir for colonization, but the mechanisms by which PA transitions from an asympt...

Microdissection techniques have become an important tool to link histomorphology and pathophysiological events using modern methods of molecular biology. They allow isolation of cell clusters or even single cells precisely under optical control from complex tissue structures for further analysis of DNA, RNA, and proteins. In particular, the fragile RNA molecules can be preserved during microdis...

A reverse transcription nested PCR (RT-PCR) assay for the detection of rubella virus RNA using primers from the E1 open reading frame was established. This assay was found to be sensitive (detecting approximately two synthetic RNA copies and RNA extracted from 0.1 50% tissue culture infective dose of rubella virus) and specific; five wild-type rubella strains and four vaccine strains were detec...

Die Aufklärung von RNA Strukturen, entweder in Isolation oder im Komplex, ist essentiell, um die Mechanismen zellulärer Prozesse zu verstehen. Festkörper-NMR auf Komplexe mit hohem Molekulargewicht anwendbar und benötigt keine Kristallisation; Methode daher gut geeignet als Teil großen mehrkomponentigen Komplexen untersuchen. Kürzlich konnten wir ersten Strukturen einem RNA-Protein Komplex mitt...

Chalcone synthase (CHS) is a plant-specific enzyme that synthesises naringenin chalcone, an essential precursor of the flavonoid biosynthetic pathway. Naringenin and kaempferol are two flavonoids that have been demonstrated to inhibit the proliferation of HeLa cells. To study chalcone synthase gene regulation in Melastoma decemfidum, we developed a high-yield total RNA isolation method to assem...