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The search result is from 1 to 4 pairs of primers. The results should be as close as possible to the amplified fragment. That means, that the searching process starts from the edge of the amplified fragment to the 5' direction on the direct strand in case of the forward primer and to the 3' direction of the reverse-complementary strand in case of the reverse primer. The results are defined with special names:

  • A - the pair of primers closest to the amplified fragment, corresponding to the entire set of parameters.
  • B1 - the pair of primers closest to the amplified fragment, corresponding to the entire set of parameters, if B2 and/or B3 exist. Could be the same as A, but not always.
  • B2 - the pair of primers corresponding to the entire set of parameters, which has a 4-nucleotide intersection with B1.
  • B3 - the pair of primers corresponding to the entire set of parameters, which starts immediately when B1 ends.

The example of a forward primers result.

Also, you may see results in the result table. Single click on the result in the result table select the corresponding annotation, double click - export the result primer to the separate file (with the backbone sequence, which will be added during the exporting process).