Touchdown (TD) PCR represents a versatile one-step procedure for optimizing PCRs even if the degree of primer-template complementarity is not fully known. The protocol relies on incremental annealing temperature decreases in progressive cycles designed to bracket the melting temperature (Tm) of the reaction. Here we investigate the characteristics of TD PCR that serve to minimize the need to op...

On-site detection by flow-through polymerase chain reaction (PCR) microfluidic systems for rapid and highly sensitive analysis, are significantly desired for bioanalytical and medical research. The conventional continuous-flow PCR chips realized rapid detection, but their sensitivity was very low (10(6) to 10(8) copies μL(-1)). We improved this drawback by coating the chip with a PCR reagents m...

Patients with end stage renal disease have the option of utilizing peritoneal dialysis (PD) as their mode replacement therapy. There are basically 2 types PD which include chronic ambulatory (CAPD) and automated dialysis. APD is most preferred type utilized globally because convenience nighttime therapy but requires a cycler machine. Unfortunately not available in Pakistan, hence utilization Pa...

Quantitative PCR (qPCR) is more sensitive than microscopy for detecting Pneumocystis jirovecii in bronchoalveolar lavage (BAL) fluid. We therefore developed a qPCR assay and compared the results with those of a routine immunofluorescence assay (IFA) and clinical data. The assay included automated DNA extraction, amplification of the mitochondrial large-subunit rRNA gene and an internal control,...

Herein we present an integrated microfluidic device capable of performing two-step gene synthesis to assemble a pool of oligonucleotides into genes with the desired coding sequence. The device comprised of two polymerase chain reactions (PCRs), temperature-controlled hydrogel valves, electromagnetic micromixer, shuttle micromixer, volume meters, and magnetic beads based solid-phase PCR purifica...

Real-time PCR at the single bacterial cell level is an indispensable tool to quantitatively reveal the heterogeneity of isogenetic cells. Conventional PCR platforms that utilize microtiter plates or PCR tubes have been widely used, but their large reaction volumes are not suited for sensitive single-cell analysis. Microfluidic devices provide high density, low volume PCR chambers, but they are ...

A point-of-care, diagnostic system incorporating a portable thermal cycler and a compact fluorescent detector for real-time, polymerase chain reaction (PCR) on disposable, plastic microfluidic reactors with relatively large reaction volume (ranging from 10 µL to 100 µL) is described. To maintain temperature uniformity and a relatively fast temperature ramping rate, the system utilizes double-si...

Pasteurella multocida (P. multocida) is an aerobic Gram-negative coccobacillus, which is found as part of the natural oral flora of many animals, including most healthy cats and dogs. However, it can cause a variety of infections in humans, usually as a result of the patient being bitten or scratched by a cat or dog. There have been 22 reported cases of P. multocida peritonitis in patients unde...

Battery thermal management is critical for high-power battery packs commonly used in electric vehicles (EV) and hybrid electric vehicles (HEV). To design battery thermal management systems properly, obtaining accurate heat generation data from battery modules is essential. To measure heat generation from full-size, multiple-cell battery modules, we developed and tested a custom-made calorimeter...