Fluorescence in situ hybridization (FISH) requires nucleic acid probes, including deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or nucleic acid analogs, labeled directly with fluorophores, or capable of indirect association with fluorophores. The nucleic acid provides the FISH assay with its specificity through complementary pairing of the probe nucleotides with nucleotides of the target...

In this thesis, we tried to improve the initiation of Rolling Circle Amplification (RCA) on genomic DNA using an approach known as Single Primer Extension (SPE). Usually, a synthetic DNA target provides a high signal on RCA after padlock probe hybridisation due to its short 3’ end. However, unfragmented genomic DNA provides low efficiency for RCA initiation due to its long 3’ end. During SPE, t...

A recombinant clone, which covers the pepsinogen gene in a single insert, has been isolated by screening a library of human genomic DNA, using a swine pepsinogen cDNA as a probe. Sequence analysis of coding DNA segments of the clone revealed that the pepsinogen gene occupies approximately 9.4-kilobase pairs of the genomic DNA and is separated into nine exons by eight introns of various lengths....

methods this experimental study was conducted on iranian children communities from may to november 2014. forty stool samples were obtained from laboratories and investigated for heat-labile toxin (lt). specific primers were designed and the dig -labeled pcr products were bounded to streptavidin-coated wells of a microtiter plate and detected by anti-dig-peroxidase conjugate. an internal biotin-...

abstract: a pure nucleotide pool is an essential precursor of correct dna replication and prevention of mutagenesis and abnormality in a living cell. inosine triphosphatase (itpase) is a critical enzyme to remove any deaminated rough purine nucleotides such as inosine from nucleotide pool. it has been shown that the rate of substitution mutation in genomic dna increase by abnormal function and ...

Gene dosage plays a critical role in a range of cellular phenotypes, yet most cellular expression systems use heterologous cDNA-based vectors which express proteins well above physiological levels. In contrast, genomic DNA expression vectors generate physiologically-relevant levels of gene expression by carrying the whole genomic DNA locus of a gene including its regulatory elements. Here we de...

The division of total genomic libraies into chromosome-specific sublibraries is critical for efficient analysis and mapping of the genome. A new colony hybridization-based approach to rapidly identify chromosome-specific clones from a human genomic bacterial artificial chromosome (BAC) library (1,2) is described. The most ideal hybridization probe for identifying all of the chromosome-specific ...

Meckel-Gruber syndrome type 3 is an autosomal recessive genetic defect caused by mutations in TMEM67 gene. In our previous study, we have identified a homozygous TMEM67 mutation in a Chinese family exhibiting clinical characteristics of MKS3, which provided a ground for further PGD procedure. Here we report the development and the first clinical application of the PGD for this MKS3 family. Mole...

Paper-based DNA biosensors are powerful tools in point-of-care diagnostics since they affordable, portable, user-friendly, rapid and robust. However, their sensitivity is not always as high required to enable quantification. To improve the response of standard dot blots, we have applied a new enhancement strategy that increases assays based on use biotinylated silica-nanoparticles (biotin-Si-NP...

We have developed a technique, called genomic subtraction, for isolating the DNA that is absent in deletion mutants. The method removes from wild-type DNA the sequences that are present in both the wild-type and the deletion mutant genomes. The DNA that corresponds to the deleted region remains. Enrichment for the deleted sequences is achieved by allowing a mixture of denatured wild-type and bi...