We present a protocol for effi cient amplifi cation of a large number of DNA targets using a single-tube polymerase chain reaction (PCR) based on PCR suppression. This method allows amplifi cation of each DNA target with only one target-specifi c primer whereas the other primer is common for all targets. Thus, this approach requires n+1 primers for n targets instead of 2n in conventional PCR an...

I n the last few years, molecular hybridization methods have been used extensively in basic and applied virology because of their technical flexibility and high specificity. Using these techniques, the detection of DNA and RNA viruses directly from clinical specimens, the analysis of the specific transcriptional activity of viral genes in vitro and in vivo, and the study of virus-host relations...

The respiratory pattern generator modulates the sympathetic outflow, the strength of which is enhanced by challenges produced by hypoxia. This coupling is due to the respiratory-modulated presympathetic neurons in the rostral ventrolateral medulla (RVLM), but the underlining electrophysiological mechanisms remain unclear. For a better understanding of the neural substrates responsible for gener...

MOTIVATION We developed a technique and a tool for degenerate primer design based on multiple local alignments employing the MEME algorithm supported with electronic PCR. The objective is to find adequate primers starting from sequences with poor global similarity. We show an example of its application in our laboratory to find sequences in Brachiaria with similarity to ESTs related to apomixis.

Inflammatory cytokines have an important role in the pathogenesis of myocarditis, but still little is known about the importance of interferon gamma (IFNg) in this disease. The aim of the study was to evaluate the prognostic value of the initial transcriptional activity of IFNg and two subunits of its receptor as measured with the use of QRT-PCR and SYBRGreen chemistry in the group of 63 patien...

A quick in vitro mutagenesis method for the construction of nested deletion libraries was developed. Many deletions can be obtained in a single inverse PCR (IPCR) by replacing one of the two primers with a mixture of 5'-truncated oligodeoxynucleotides. Since chemical DNA synthesis proceeds from the 3'to the 5'end, such a mixture of 5'-truncated oligodeoxynucleotides can easily be obtained in a ...

were seropositive. Because antibodies may be detectable 10 days–3 weeks after experimental infection for the first time (8), the presumed period of infection was between midJanuary and mid-February. At this time, the highest temperatures again rose above 6°C for a few days (Figure 1, panel A). Although the within-herd seroprevalence was >90% in ewes after confirmed or suspected SBV infection in...

Figure A2. QRT-PCR analysis of OR expression for OR families 4, 5, 7, and 9. There was no statistical difference of OR expression between PEA (red bars) and control fish (blue bars) for any of the families tested. Similar results were observed for two different stocks (#1111 (dark shades) and #1106 (light shades)) of imprinted fish. All OR expression data was normalized to expression of β-actin .

To the editor: We wish to offer some cautionary remarks concerning the report by de Graaf et al. [1] about the first human tick-borne encephalitis (TBE) infection in the Netherlands, acquired in June 2016 (and not in July, as incorrectly mentioned in the title). At first sight, this case, apparently proven by ELISA and confirmed by neutralisation tests (NT), seems extremely convincing, especial...

While many data are available on genes encoding proteins for degradation of hydrocarbons in bacteria, the impact of alkane on transporter protein expression is unclear. Pseudomonas aeruginosa SJTD-1 is a strain that can consume medium- and long-chain n-alkanes. In order to study the proteins involved in n-octadecane uptake, we use iTRAQ and label free comparative proteomics analysis to identify...