UNLABELLED PREMISE OF THE STUDY Development of genetic markers can be costly and time-consuming, especially when multiple primer pairs are fluorescently labeled. This step was streamlined by combining two techniques in the same PCR reaction: (1) custom-labeling of primers by the investigator and (2) multiplexing multiple primers together in the same reaction. • METHODS AND RESULTS This tec...

Acidic fibroblast growth factor (aFGF) is a heparin binding protein that displays pleiotropic activity. The purpose of this study was to document the presence of the translated aFGF product, its mRNA, and its location in the colon. mRNA was extracted from bovine large intestine and reverse transcribed to cDNA. Nested-primer PCR was used to determine the presence of mRNA using primers homologous...

A nested polymerase chain reaction (PCR) was developed to detect genomic DNA of Piscirickettsia salmonis, the causative agent of an epizootic disease in salmonids. The nested PCR assay, which used general bacterial 16s rDNA primers in the flrst amplification reaction, and P salmonisspecif~c primers in a second reaction, allowed detection of less than 1 P salmonis tissue culture infect~ous dose ...

Genus and Species. Bos indicus and Bos taurus. Locus Name. Bovine stearoyl-coenzyme A desaturase (SCD). Source and Description of Primers. Primers were designed that amplified a 135-bp region in SCD exon 6. Sequences from human (GenBank accession number Y13647), rat (AB003357), mouse (M21285), and pig (Z97186) were aligned to identify a conserved region in which to design the primers. The forwa...

Current methods to detect hepatopancreatic parvovirus (HPV) infection of penaeid shrimp depend on invasive techniques that require dissecting the organs infected by this virus. However, sacrificing valuable stocks in order to determine their HPV status can be a drawback in the case of breeding programs. A method was developed for HPV detection by applying a polymerase chain reaction (PCR) assay...

DNA family shuffling is a powerful method for enzyme engineering, which utilizes recombination of naturally occurring functional diversity to accelerate laboratory-directed evolution. However, the use of this technique has been hindered by the scarcity of family genes with the required level of sequence identity in the genome database. We describe here a strategy for collecting metagenomic homo...

Methods Wild type human von Willebrand factor cDNA: Von Willebrand factor cDNA was cloned from the Marathon-ready human bone marrow cDNA library (Clontech, Mountain View, CA) using the following three sets of primers: 5'-CCATCCTAATACGACTCACTATAGGGC-3’ / 5'TCAATCTCTCCTCCCTCCACCAGGAT-3' (VWF 3-3103), 5'CTCACCTTCGACGGGCTCAAATACCTG-3' / 5'-TGGGAGCTGTCGGGGACAAGACAC-3' (VWF 2925-6712), 5'-AGCGAATCGAA...