Primer length:Oligonucleotides between 18-24 are said to be quiet enough and advantageous so that short primers would bind easily to the template at the annealing temperature. Melting temperature (52°C-56°C)The GC results of the sequence gives a fair indication of the primer Tm. However, t...
It is important, however, to maintain fairly tight control over the melting temperature range of the primers involved. One worthwhile strategy is to prepare separate pools of primers, the members of each of which exhibit a similar Tm. Of course, it is also possible to use only one primer,...
(Wittwer, 2009). Using slow denaturation of the PCR product after the final amplification cycle leads to gradual denaturation of the double-stranded DNA. The melting temperature, defined as the temperature at which half of the DNA is denatured, depends on the composition and the length of the ...
The software calculates melting temperature (Tm) of oligos using the "nearest neighbor" algorithm. Documents for The Primer Express software also provides documents for the following Sequencing sequencing applications: o Cycle Sequencing o Sequencing Primer Batch Processing For speed and convenience, the ...
42、ssion numbers, definition, or sequence length by clicking the respective column headers. You can also delete the selected sequence(s).Search Status TabFor convenience, an overall view of the designed primer is given in the Search Status Tab at the end of each search.The search status is...
2. Customize at least 1 criterion (Table 1) used to design your primers and probe. Hints: Click on any criterion name to open a window with a definition of the criterion. If needed, select aDesign Parameterradio button again to reset all the criteria back to default settings. ...
Our tool ensures most convenient primer design, because it only requires the absolute minimum of parameter definition. This is mainly the genome position to be addressed which is easily accessible through GenBank or SimpleSearch or even already known. In addition, the distance to the target ...
*Definition of CT value The CT (cycle threshold) value of a real-time PCR procedure is the number of cycles required for the fluorescent signal to exceed the background level i.e. "cross the threshold". CT values are dependent on various factors including primer design and DNA template conc...
LNA probes could be made shorter than the DNA counterparts, while still maintaining high melting temperature to increase specificity. The LNA probes clearly outperformed DNA probes to give signal only when a completely matched target was present. In general, the unique hybridization capability observed...
“specific region” of a primer depends mainly on the averaged lengths of the predicted PCR products based on bioinformatic calculations. The definition includes, without limitation, a “specific region” of between about 4 and 12 bases in length. In more specific embodiments, the length of the ...