Gene expression analysis has become an invaluable tool for understanding gene function and regulation. However, global expression analysis requires large RNA quantities or RNA preamplification. We describe an isothermal messenger RNA (mRNA) amplification method, Ribo-SPIA, which generates micrograms of labeled cDNA from 5 ng of total RNA in 1 day for analysis on arrays or by PCR quantification....

We have postulated that the catecholamine-synthesizing enzymes tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), and phenylethanolamine N-methyltransferase (PNMT) are coded for by similar genes. To analyze the structural relationship of genes coding for these enzymes, we have cloned DNAs complementary (cDNA) to DBH and PNMT messenger RNAs (mRNAs). Using hybrid selection analysis to id...

Introduction The possibility to amplify total cDNA obtained from small amounts of biological material is not yet routinely considered, despite the fact that obtaining amounts of material suitable for direct processing by standard methods is often time-consuming, expensive and may be even impossible. Perhaps the most significant obstacle to the full appreciation of the technique is the widesprea...

Background: Breast cancer is one of the most prevalent malignancies among women in various countries. HER2 overexpression, which is due to different reasons, occurs in 20-30% of breast cancers. HER2 gene encodes an 185kDa transmembrane glycoprotein with 1255 amino acids. This active product triggers downstream intracellular signaling pathways inducing cell proliferation and cell survival. These...

backgrounds and aims: coxsakievirus b3 (cvb3), one of the six coxsakievirus b serotypes, is a member of the enterovirus genus within the picornaviridae family. cvb3 is an important pathogen of viral myocarditis, which accounts for more than 50% of viral myocarditis cases. the genome of cvb3, like that of other entroviruses, is a single-stranded, sense, polyadenylated rna molecule with 7400 nucl...

Name of the Sequence. Red seabream β-actin cDNA. Genus and Species. Pagrus major. Origin of Clone. Total RNA was isolated from ovary of red seabream. Poly(A)+ RNA was purified using an Oligotex-dT30 (Super) mRNA purification kit (Takara, Japan). Double-stranded complementary DNA synthesis was carried out using a TimeSaver cDNA synthesis kit (Pharmacia Biotech, Uppsala, Sweden). The cDNA library...

Several site-directed mutagenesis methods have been developed and are being currently used in research. All these methods apply three or four primers, including one or two bearing the appropriate mutation, and most use PCR to amplify the desired sequence and to incorporate the mutation (1–6). Although several developments have improved the robustness and reliability of the method, all of them r...

A label-free colorimetric method based on exonuclease I (Exo I)-assisted signal amplification with protamine as a medium was developed for analysis of kanamycin. In this study, double-stranded DNA (dsDNA) probe tailored by manipulating an aptamer and its complementary (cDNA) ensuring detection target high selectivity excellent sensitivity. Herein, could not only combine negatively charged gold ...

secreted cytokines of th1 (t-helper)/th2 cells play an important role in the pathogenesis of many diseases. th1 cells secrete predominantly ifn-γ and il-2 which regulate cell-mediated immunity against intracellular pathogens and tumors. in this study, expression of ifn-γ was studied using semiquantitative rt-pcr. in brief, lymphocytes of a healthy donor were stimulated with pha (1μg/106 cell/ml...