After sample processing for RNA and DNA analysis, the leftover protein pellets are usually discarded due to limited efficiency of pellet reconstitution/solubilisation. As pelleted proteins tightly packed, they most often solubilised using chaotropic agents (e.g., guanidine hydrochloride or urea), detergents SDS), salts (NaCl) basic buffer (Tris). The aim this study was define optimise procedure...

OBJECTIVE OF WORK The aim of this study was to compare three methods of RNA extraction for molecular analysis of oral cytology to establish the best technique, considering its concentration and purity for molecular tests of oral lesions such as real-time reverse transcriptase reaction. MATERIAL AND METHODS The sample included exfoliative cytology from the oral cavity mucosa of patients with n...

BACKGROUND According to mRNA microarray, proteomics and other studies, biological abnormalities of eutopic endometrium (EM) are involved in the pathogenesis of endometriosis, but the relationship between mRNA and protein expression in EM is not clear. We tested for the first time the hypothesis that EM TRIzol extraction allows proteomic Surface Enhanced Laser Desorption/Ionisation Time-of-Fligh...

Complete inactivation of infectious Ebola virus (EBOV) is required before a sample may be removed from a Biosafety Level 4 laboratory. The United States Federal Select Agent Program regulations require that procedures used to demonstrate chemical inactivation must be validated in-house to confirm complete inactivation. The objective of this study was to develop a method for validating chemical ...

doi: 10.1111/j.1399-3054.2006.00716.x Highly sensitive techniques for transcriptome analysis, such as microarrays, complementary DNA-amplified fragment length polymorphisms (cDNAAFLPs), and others currently used in functional genomics require a high RNA quality and integrity, as well as reproducibility among extractions of replicates from the same tissue. There are, however, few technical paper...

The main objective of this study was to isolate high-quality total ribonucleic acid (RNA) from raw fresh semen and frozen-thawed boar semen, using a protocol comprising the conventional TRIzol assay and a membrane-based technique, the PureLink RNA mini kit. Bioanalyzer profile revealed that the sperm RNA size distributions comprised mainly intact RNA ranging from 1500 to 1800 bp, without any de...

To the Editor: Preservation of RNA integrity is important in microarray techniques for identifying differentially expressed genes so that results reflect true biological differences and not differences in RNA degradation (1 ). RNA degradation is usually low in RNA isolated from cultured cells (2 ). When samples isolated from RNaserich tissues are used, however, RNA degradation may introduce bia...

The extraction of high-quality ribonucleic acid (RNA) from tissues and cells is a key step in many biological assays. Guanidinium thiocyanate-phenol-chloroform (AGPC) widely used efficient method to obtain pure RNA most cells. However, it not with some like sperm because they are resistant chaotropic lysis solutions containing guanidinium thiocyanate such as Buffer RLT+ Trizol. Here, we show th...

Piotr Chomczynski Department of Internal Medicine School of Medicine University of Cincinnati Cincinnati, Ohio 45267 Progress in many fields of basic and clinical research depends on the availability of high-quality RNA. Of the many methods for isolating RNA (reviewed in 1 and 2), a single-step RNA isolation method based on guanidine isothiocyanate/phenol/chloroform extraction (3) has been grow...