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The search can be done into the target sequence also by clicking the appropriate radio buttion. One can then copy the current selection and paste it either directly or by this menu in a new line of the primer list.

Although its first finality is to test already existing primers quickly, AmplifX incorporates since version 1.

A new window opens which makes it possible to fix some specific parameters to this function, such as the length of the primers wished, the difference of TM between the two ones and the minimum score of quality being used as threshold.

This calculation of quality is also dependent on certain parameters which can be changed by the menu " Preferences", such as for example, TM or GC composition.

Once the search is finished, one can send the primers selected by double-clicking onto the list and clicking on the button "Add to primer list".

Tip : Do not specify a large searching zone for each primer restricting to bp is reasonable , and start by fixing the quality score threshold to a high value arround If no primer pairs were found, decrease the constraints then.

If you want to calculate primers in more sophisticated way, I advise you to use a specialized application such as Primer3 [Steve Rozen and Helen J.

Skaletsky Primer3 on the WWW for general users and for biologist programmers. Humana Press, Totowa, NJ, pp ] , a good implementation of this program can be found at this address.

Skip to main content. Manage, test and design primers for PCR And do more with version 2! What is AmplifX? Many algorithms has been rethought in regard to new insights and real experimental designs in the field of PCR.

Many new features and bug corrections! Amplification with primers that overhangs the target, opening an older AmplifX version regarding of the pref file [2.

The algorithm starts seeking perfect matches of a sub-sequence seed in terminal position 3' of the primer against the whole target sequence and its complementary sequence.

For each position where the seed matchs, a "priming score" is calculated based onto data from Stadhouders et al. If the score is above an user-modifiable cutoff, the match is kept.

If N is higher than the seed size then it means that one has a chance to get a primer that has at least one mismatch to the target at any position i.

Primers with TM and GC comprised between the min and max cutoffs are now considered all as good. For long primers the calculation is done only for the 3' moiety judged relevant primer with a 5' flanking sequence designed for sequencing or cloning, etc.

The user can mark this by using different font cases lower or upper in the sequence. In absence of such case-marked regions the calculation uses only the last 25 nucleotides.

On the contrary differences of cases in the last 3' 17 nucleotides are not taken into account. The calculation does not use the previous "polyX" parameter but computes now a lexical complexity score that is more powerful and discriminative.

Improvements of the sequence and primer drawings. The zoom can now be at the nucleotide level displaying in letters. The zoom centers inside the selection Improvement of the zoom management.

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