Traditional RNA extraction methods rely on the use of hazardous chemicals such as phenol, chloroform, guanidinium thiocyanate to disrupt cells and inactivate RNAse simultaneously. RNA isolation from Caenorhabditis elegans presents another challenge due to its tough cuticle, therefore several repeated freeze-thaw cycles may be needed to disrupt the cuticle before the cell contents are released. ...

during 2009-10, real time rt-pcr and conventional rt-pcr techniques used for detecting btvs rna in 310 blood samples. for real time and gel based rt-pcr segment-1 and segment-10 selected as conserve genes to search any btv strains. using these methods, 58 (%18.7) and 14 (%4.5) positive samples were detected among the clinically suspected sheep. sensitivity of both molecular techniques evalua...

Poliovirus-specific double-stranded RNA molecules containing covalently attached protein were coupled with a biotin ester through the protein moiety. Subsequent interaction of the RNA-biotin with avidin attached to electronopaque plastic spheres led to the formation of complexes that were easily visualized in the electron microscope. Avidinspheres were associated only with one end of the RNA-bi...

RNA extraction from mangrove tissues can be difficult due to the presence of polyphenolic and polysaccharide compounds upon cell disruption. Besides, a successful RNA isolation from mangrove tissues sometimes can be strain and species specific. Two different methods were used to extract RNA from tissues (stems, leaves and roots) of black mangrove (Avicennia germinans) and white mangrove (Lagunc...

background: rna interference (rnai) is a phenomenon uses double-stranded rna (dsrna) to specifically inhibit gene expression. the non-specific silencing caused by interferon response to dsrna in mammalian cells limits the potential of utilizing rnai to study gene function. duplexes of 21-nucleotide short interfering dsrna (sirna) inhibit gene expression by rnai. in some organisms, sirna can als...

Laser-capture microdissection and transcriptional profiling have enabled compartment- and cell-specific analysis of gene expression in chronic kidney disease, thus facilitating the investigation of pathophysiological associations between glomerular, tubular, and interstitial structures. Due to the pico- and nanogram amounts of RNA isolated from LCM-captured material linear RNA amplification pro...

RNA-binding proteins (RBPs) are key mediators of posttranscriptional gene expression control. However, the links between cell signaling on one hand and RBP function other understudied. While thousands posttranslational modification (PTM) sites RBPs have been identified, their functional roles only poorly characterized. RNA-interactome capture (RIC) cross-linking immunoprecipitation (CLIP) attra...

We have developed a procedure for isolation of microRNA and genomic DNA in addition to total RNA from whole blood stabilized in PAXgene Blood RNA tubes. The procedure is based on automatic extraction on a BioRobot MDx and includes isolation of DNA from a fraction of the stabilized blood and recovery of small RNA species that are otherwise lost. The procedure presented here is suitable for large...

Isolation of proteins that specifically interact with a given RNA or RNA regulation element is essential for studies on the molecular mechanisms of gene expression. Here, a novel method for direct isolation of such interacting proteins is described. It uses an affinity medium that consists of an interacting RNA with an artificially added 'tail', which is annealed to one end of a DNA 'arm', the ...