The use and validation of a strategy that allows a universal set of bar-coded sequencing primers to be appended to an amplified PCR product is described. The strategy allows a modular approach, in that the same bar code can be used with two or more target-specific primer sets, even simultaneously.

In the present study, we developed a quick, highly specific method for detection of Shigella species by combining immunocapturing of the bacteria and a universal primer PCR. The method drastically enhances test sensitivity, and it can be used not only for identification of Shigella species in the environment but also for rapid detection of other pathogens.

Novel methods were developed to isothermally detect target nucleic acids and initiate signal amplification cascades. The methods utilize target-specific MNAzymes to activate universal primer molecules. These primers can subsequently promote the autonomous synthesis of DNAzymes capable of cleaving nucleic acid substrates and generating signal, which further creates the potential for circular fee...

The diversity of protozoan-associated methanogens in cattle was investigated using five universal archaeal small-subunit (SSU) rRNA gene primer sets. Methanobrevibacter spp. and rumen cluster C (distantly related to Thermoplasma spp.) were predominant. Significant differences in species composition among libraries indicate that some primers used previously to characterize rumen methanogens exhi...

A collaborative study initiated from the first annual European Union (EU) Concerted Action Group meeting (project PL 98-4032, 5-7 May 1999, Federal Research Centre for Virus Diseases of Animals, Tübingen, Germany) was organised between the Veterinary and Agrochemical Research Centre in Brussels, the Federal Research Centre in Tübingen and the OIE/FAO World Reference Laboratory for Foot-and-Mout...

Polymerase chain reaction (PCR) as a method for preparing DNA templates has been used for several DNA sequencing applications. An in situ procedure for directly sequencing PCR products by the dideoxy-termination method has been developed by using fluorophore-labeled sequencing primers. Completed sequence reactions were combined and loaded into a single electrophoretic lane of a fluorescence-bas...

Several methods for the detection of RNA have been developed over time. For small RNA detection, a stem-loop reverse primer-based protocol relying on TaqMan RT-PCR has been described. This protocol requires an individual specific TaqMan probe for each target RNA and, hence, is highly cost-intensive for experiments with small sample sizes or large numbers of different samples. We describe a univ...

BACKGROUND Plasmid encoded blaCTX-M enzymes represent an important sub-group of class A beta-lactamases causing the ESBL phenotype which is increasingly found in Enterobacteriaceae including Klebsiella spp. Molecular typing of clinical ESBL-isolates has become more and more important for prevention of the dissemination of ESBL-producers among nosocomial environment. METHODS Multiple displacem...

The caption for Figure 5 reads: One-color SNV quantification. (A) Primers are designed with the single nucleotide variant at the 3′ end of the complementary region. Noncomplementary tails of varying lengths are then added to the 5′ end and amplified with a universal reverse primer. (B) 1:4 mixture of MUT/WT BRAF template amplified with mutant primers with the short tail and wild-type primers wi...