Detection of RNA viruses by reverse transcription (RT)-PCR has proven to be a useful approach for the diagnosis of infections caused by many viral pathogens. However, adequate controls are required for each step of the RT-PCR protocol to ensure the accuracies of diagnostic test results. Heterologous competitor RNA can be used as a control for a number of different aspects of diagnostic RT-PCR. ...

Gene expression studies in equine research involve the use of whole blood samples as a vital source of RNA. To determine the optimal method for RNA isolation from equine whole blood, we compared three RNA isolation strategies using different commercially available kits to evaluate the yield and quality of equine RNA. All 3 methods produced RNA with high quality. Though it did not produce the hi...

RNA isolation is essential to the study of gene expression at the molecular level. However, it is difficult to isolate RNA from organisms that contain large amounts of polysaccharides or other compounds that bind or coprecipitate with RNA, such as the unicellular protist Euglena gracilis. Currently, there is no commercial kit available that is specific for the isolation of high-quality RNA from...

Current RNA isolation methods have limitations in their ability to yield good quality and quantity of RNA from plants that have high content of phenols, polysaccharides and storage proteins. Existing methods also do not eliminate accompanying chromosomal DNA in RNA preparation that causes false positives in gene expression studies. Standard isolation technique was modified for rapid and quick e...

background: recently, the use of t7 rna polymerase instead of other viral and cellular promoters is increasing due to high efficacy of transcription in the cell cytoplasm by this polymerase. in order to translate the transcripts produced by t7 rna polymerase in mammalian cell lines, it is necessary to include internal ribosome entry site (ires) sequences. in addition, if sequence of poly a sign...

RNA quality is of utmost importance to perform gene expression quantification by qPCR. The classical methods used to determine RNA quality are based on electrophoresis and spectrophotometer assessment, namely A(260)/A(280) and A(260)/A(230) ratios. It was previously shown that due to the complex nature of Staphylococcus epidermidis biofilms, RNA extraction procedures could impact mRNA quality a...

L-cells were fractionated at various times after infection with EMC virus and assayed for haemagglutinin, virus and phenol-extractable infective RNA. A second fraction of infective RNA was obtained by sodium dodecyl sulphate (SDS) extraction of the samples after phenol extraction. The infectivity of SDS-extracted RNA increased earlier than that of phenol-extracted RNA, but the maximum titre was...

There are several protocols and kits for the extraction of circulating RNAs from plasma with a following quantification of specific genes via RT-qPCR. Due to the marginal amount of cell-free RNA in plasma samples, the total RNA yield is insufficient to perform Next-Generation Sequencing (NGS), the state-of-the-art technology in massive parallel sequencing that enables a comprehensive characteri...