Simplified method for confirmation of PCR products

Simplified method for ligase-free cloning of PCR products.

Cloning of polymerase chain reaction (PCR) products into plasmid vectors by conventional methods, relying either on recognition sequences built into the primers or taking advantage of the template-independent terminal transferase activity of Taq DNA polymerase to add an extra adenine to the 3′ end of the product, are often difficult (1,10,12). In recent years, numerous cloning strategies (10,13...

An efficient method for purification of PCR products for sequencing.

A high-throughput DNA sequencing method that generated high quality data was developed. A frame fashioned from a standard agarose gel combined with 0.1%-0.2% low-melting point (LMP) agarose gel was used to isolate the PCR product of interest. Collected PCR products were centrifuged without any reagents and the supernatants were directly used for a sequencing reaction. This method is simple and ...

Evaluation of synthetic DNA probes for confirmation of Clostridium perfringens enterotoxin gene PCR products.

A new diagnostic reagent was developed that is capable of detecting the presence of Clostridium perfringens rapidly and accurately compared to the conventional methods. C. perfringens enterotoxin (cpe) gene is the gene of interest since it encodes the enterotoxin responsible for food poisoning. Two new cpe-specific labeled DNA probes were evaluated using Southern and dot blot hybridization. Bac...

PCR confirmation of infection with Cyclospora cayetanensis.

persons and the public health relevance of this emerging pathogen as a potential cause of diarrheal outbreaks (3,4) make prompt disclosure of the epidemiologic features and behavior of the parasite necessary. As we propose the possible participation of poultry in the epidemiologic cycle of the coccidia, we invite other Cyclospora working groups worldwide to confirm the so far putative reservoir...

A simplified method for PCR detection of hepatitis C virus RNA from human serum.