The bovine serum albumin (BSA) protein is employed in genosensors as nonspecific binding blocker. In this work, the influence of BSA on electrochemical response a DNA-based biosensor explored by impedance spectroscopy (EIS) and double pulse voltammetry technique. Each buffer component hybridization studied bare gold electrodes single-stranded DNA (ssDNA)-functionalized electrodes. found to have...

Chitinase cDNAs from Leucaena leucocephala seed]ings were cloned by PCR amplification with degenerate primers based on conseryed class I chitinase sequences and cDNA library screening. Two closely related chitinase cDNAs were sequenced and inferred to encode precursor proteins of 323 (KBI) and 326 (KB2) amino acids. Expression of the KB2 chitinase from a pET32a plasmid in Origami (DE3) Escheric...

The accurate evaluation of telomerase activity, a typical cancer biomarker, is vital for early screening. In this study, we developed dark-field microscopy (DFM) visual single-particle detection scheme to detect activity based on automatic counting gold nanoparticles (AuNPs). This method started with attaching the substrate (TS) primer magnetic beads (MBs) through streptavidin-biotin interactio...

Gene expression profiling is the characterization of cells based on the level of gene activity represented by concentrations of complementary DNA reverse transcribed from messenger RNA. The spectrum of cDNA concentrations, the expression profile, is determined using a DNA microarray. Although this approach is valuable for research, a simpler scheme that would give answers on a shorter time-scal...

background: because of the strong immunologic responses of surface protein tasp in theileria annu­lata infected host, we tried to characterize this protein in a t. annulata isolate from iran. methods: the rna prepared from t. annulata infected cells was used to produce smart-ds-cdna. the double strand cdna was then amplified with primers derived from tasp mrna se­quences. the pcr product was cl...

A LEAFY cDNA was cloned from the short-day (SD) plant Brassica juncea cv Varuna. (BjLFY) consists of 1261 bp encoding for a protein of 420 amino acids and an estimated isoelectric point of 6.8. The deduced amino acid showed 99% and 86% identity to Arabidopsis thaliana and Brassica oleracea LFY cDNA respectively. The LFY transcript was detected throughout the vegetative and reproductive phase bu...

Full-length cDNA (FLcDNA) libraries consisting of 172,000 clones were constructed from a two-row malting barley cultivar (Hordeum vulgare 'Haruna Nijo') under normal and stressed conditions. After sequencing the clones from both ends and clustering the sequences, a total of 24,783 complete sequences were produced. By removing duplicates between these and publicly available sequences, 22,651 rep...

Obtaining full-length cDNA is important for many molecular biology methods like cDNA library construction or RACE-PCR (rapid amplification of cDNA ends). We have found that the inclusion of betaine alone and in combination with trehalose in reverse transcription results in longer cDNA synthesis products. As shown on the 14-kb long clarithrin mRNA with real-time PCR, a combination of 2 M betaine...

Using as criteria the size, abundance and developmental specificity of hybridizing mRNA sequences, we have selected from our chorion cDNA library a clone corresponding to a specific chorion protein, A4--cl. Comparison between the clone sequence and the largely known sequence of A4--cl validates the use of the cDNA library for sequence analysis of the chorion multigene families. The two major ch...

The SV4O-immortalized mouse macrophage cell line, BAC1 .2F5, is strictly dependent on CSF-1 for its survival and proliferation in culture. Introduction of a retroviral vector containing a 1 .6 kilobase (kb) pair human CSF-1 cDNA into these cells abrogated their growth factor dependence but did not render the cells tumorigenic in nude mice. The infected macrophages contained multiple copies of t...