Design of primer
In the present study, a set of 16 primers were designed
[Table 1] to amplify 14 fragments [Table 2] from
the two universal vectors pUC57 and pET21a and
subsequently amplicons used as a DNA ladder. In
this report, appropriate primers were designed by the oligo v.5 software. In order to achieve the specific
amplification, we tried to avoid formation of bothering
structures: Mismatching, mispriming, hairpin
formation and primer-primer dimmers. Primers
were selected with average length of 12-18 bp with
annealing temperature between 50ºC and 60ºC and
the 3’ ΔG above −7.8.