Quantification of human DNA has been an integral part of forensic DNA analysis. Hybridizationbased methods such as Quantiblot® kits were used extensively in the 1990s. These methods fulfilled the need at the time, since their sensitivity range was similar to the genotyping methods in use, such as restricted fragment length polymorphism. Later, the development of robust and more sensitive megapl...

AIM To apply massively parallel and clonal sequencing (next generation sequencing or NGS) to the analysis of forensic mixed samples. METHODS A duplex polymerase chain reaction (PCR) assay targeting the mitochondrial DNA (mtDNA) hypervariable regions I/II (HVI/HVII) was developed for NGS analysis on the Roche 454 GS Junior instrument. Eight sets of multiplex identifier-tagged 454 fusion primer...

introduction: listeria monocytogenes is a gram positive bacteria, frequently found in the environment and is responsible for food-borne disease such as perinatal infections, septicameia and meningeonceohalitis in human and animals. material and methods: for this reason, distribution of the ctpa determinate among l. monocytogenes isolated from clinical environment, diry and poultry samples were ...

Massively parallel sequencing of cell-free, maternal plasma DNA was recently demonstrated to be a safe and effective screening method for fetal chromosomal aneuploidies. Here, we report an improved sequencing method achieving significantly increased throughput and decreased cost by replacing laborious sequencing library preparation steps with PCR employing a single primer pair designed to ampli...

Quantitative analysis of next-generation sequencing (NGS) data requires discriminating duplicate reads generated by PCR from identical molecules that are of unique origin. Typically, PCR duplicates are identified as sequence reads that align to the same genomic coordinates using reference-based alignment. However, identical molecules can be independently generated during library preparation. Mi...

Two paradigm shifts in DNA sequencing technologies-from bulk to single molecules and from optical to electrical detection-are expected to realize label-free, low-cost DNA sequencing that does not require PCR amplification. It will lead to development of high-throughput third-generation sequencing technologies for personalized medicine. Although nanopore devices have been proposed as third-gener...

ObjectivesUndetectable or low-level hepatitis B virus (HBV) DNA and drug resistance mutations in patients may increase the risk of HBV transmission cause active viral replication other clinical problems. Here, we established a highly sensitive practical method for detection using polymerase chain reaction (PCR) -based CRISPR-Cas13a system (referred to as PCR-CRISPR) evaluated its capability sam...

background: serological assay based on dense granular (gra) proteins of toxoplasma gondii (t. gondii) is actually the most popular laboratory diagnostic tool to detection of toxoplasmosis. we aimed to construct a recombinant gra7-ptz57rt plasmid vectors that it is suitable for sub-cloning and gra7 protein production. m aterials and methods: souris mice were used for maintaining of t. gondii tac...

Bisulfite sequencing is a widely used method for measuring DNA methylation in eukaryotic genomes. The assay provides single-base pair resolution and, given sufficient sequencing depth, its quantitative accuracy is excellent. High-throughput sequencing of bisulfite-converted DNA can be applied either genome wide or targeted to a defined set of genomic loci (e.g. using locus-specific PCR primers ...

Genetic diversity as an important marker of the ecological status of aquatic ecosystems is considered a unique and powerful tool to evaluate biological communities. In order to evaluate the genetic diversity among golden mullet species (Liza aurata) in the southeast and southwest coasts of the Caspian Sea by D-Loop gene sequencing, a total of 23 fin specimens of golden mullet were collected fro...