Bovine viral diarrhoea virus (BVDV) is an important pathogen of dairy cattle. In this study, bulk milk samples representing a total of 4105 milking cows, from 18 dairy cattle herds in the suburb of Mashhad- Iran, were tested for presence of BVDV by the use of a nested reverse transcription polymerase chain reaction (Nested RT- PCR) assay. Non of the cows in the herds had been vaccinated against...

1. Besides miR-133a, miR-133b and miR-208a, there are several other muscle-specific miRNAs, such as miR-1, etc. Why only these miRNAs were used in this study? 2. P2: qRT-PCR is not quantitative real-time polymerase chain reaction, but quantitative reverse transcription-polymerase chain reaction. 3. P6: “To date, no housekeeping miRNAs have been established and validated to normalize for the miR...

In vitro produced embryos, due to their lower developmental potential when compared to the in vivo derived counterparts, have been currently subjected to an intensive scientific investigation. Qualitative as well as quantitative analyses of gene expression (reverse transcription-polymerase chain reaction; RT-PCR) belong to the powerful tools providing a wide spectrum of data on the quality of o...

In this study, 331 samples from calves less than one month old from a dairy herd in the district of Piracanjuba, state of Goiás, Brazil were tested for rotavirus. Thirty-three samples (9.9%) tested positive for rotavirus. Out of those, 31 were submitted to G and P characterization by reverse transcription followed by semi-nested polymerase chain reaction. Two samples were characterized as G6P[1...

Extensive detail on the application of the real-time quantitative polymerase chain reaction (QPCR) for the analysis of gene expression is provided in this unit. The protocols are designed for high-throughput, 384-well-format instruments, such as the Applied Biosystems 7900HT, but may be modified to suit any real-time PCR instrument. QPCR primer and probe design and validation are discussed, and...

The use of reverse transcription polymerase chain reaction (RT-PCR) with internal RNA competitive standards (competitors) provides a means for measuring absolute amounts of mRNA transcripts in small numbers of cells. Most quantitative competitive (QC)-RT-PCR methods require analysis of multiple reactions to determine the equimolar point of the products produced from mRNA vs. competitor RNA. Her...

Combining the patch-clamp method with single-cell reverse transcription polymerase chain reaction (scRT-PCR) a fusicoccin-induced current reflecting the activity of the plasma membrane H(+) ATPase of lily pollen protoplasts was measured and subsequently, the ATPase-encoding mRNAs were collected and amplified. Southern blot signals were observed in all 'patch-catch' experiments and could be dete...