Sequencing of Buchnera amino acid genes Amplification primers (table S2) were designed using the sequenced genome of Buchnera APS (NC_002528.1) and synthesized by Eurofins-MWG Operon. PCR conditions were an initial denaturing step at 94° C for 2 min, followed by 30 cycles of 94° C for 30 s, 54-60o C depending on primer Tm for 30 s, 72° C for 2-4 min depending on amplicon length, and a final ext...

This protocol describes a series of procedures for isolating nucleic acids (DNA and RNA) and proteins from moss (Physcomitrella patens) tissue. This series includes a rapid, small-scale procedure for isolating DNA, which results in genomic DNA that is only suitable for polymerase chain reaction (PCR), as well as a method for obtaining much larger amounts of genomic DNA suitable for Southern ana...

The purpose of this study was to compare the effectiveness of the QIAGEN QIAamp Stool Mini Kit against a standard phenolchloroform procedure for the extraction, quantitation, and STR-typing of human nuclear DNA from human feces. Stools from six subjects were sampled by swabbing and excision. Samples extracted with the QIAamp kit gave a wide range of DNA yields, whereas those extracted by the or...

BACKGROUND Retrograde intubation has been accepted internationally as a viable alternative for managing the difficult airway. Various techniques have been described to perform this procedure, however, difficulties have arisen on account of problems with suboptimal materials. We therefore describe a retrograde intubation technique using the knife and stiff plastic introducer from a Mini-Trach II...

Identification of Fusarium species by traditional methods requires specific skill and experience and there is an increased interest for new molecular methods for identification and quantification of Fusarium from food and feed samples. Real-time PCR with probe technology (Taqman) can be used for the identification and quantification of several species of Fusarium from cereal grain samples. Ther...

The application of high-throughput, next-generation sequencing technologies has greatly improved our understanding of the human oral microbiome. While deciphering this diverse microbial community using such approaches is more accurate than traditional culture-based methods, experimental bias introduced during critical steps such as DNA extraction may compromise the results obtained. Here, we sy...

AIM To examine the sensitivity and accuracy of real-time polymerase chain reaction (PCR) for the quantification of hepatitis B virus (HBV) DNA in semen. METHODS Hepatitis B viral DNA was isolated from HBV carriers' semen and sera using phenol extraction method and QIAamp DNA blood mini kit (Qiagen, Germany). HBV DNA was detected by conventional PCR and quantified by TaqMan technology-based re...

Isolating RNA from insects is becoming increasingly important in molecular entomology. Four methods including three commercial kits RNeasy Mini Kit (Qiagen), SV Total RNA isolation system (Promega), TRIzol reagent (Invitrogen), and a cetyl trimethylammonium bromide (CTAB)-based method were compared regarding their ability to isolate RNA from whole-body larvae of Thaumatotibia leucotreta (Meyric...

For microarray analysis, cells were seeded in 6-well plates (Corning, NY, USA) at 1×10 cells per well, and after 24 h, they were then treated with (0.0005 μg/ml) the drug either as a solution or encapsulated in nanoparticles for 5 days. Total RNA used for the microarray analysis was isolated from cultured cells using TRIZOL reagent (Invitrogen, USA) and purified using an RNeasy Mini Kit (Qiagen...

BACKGROUND Mycobacterium bovis is the aetiological agent of bovine tuberculosis (bTB), an important recrudescent zoonosis, significantly increasing in British herds in recent years. Wildlife reservoirs have been identified for this disease but the mode of transmission to cattle remains unclear. There is evidence that viable M. bovis cells can survive in soil and faeces for over a year. METHOD...