u00b0 c 15 sec 72 u00b0 c 1 5 min 65 u00b0 c 2 min plasmid detection by qpcr the number of plasmid molecules captured on the dynabeads was quantified using quantitative real time pcr amplification of a short amplicon from the template plasmid was detected by measuring the fluorescence generated from incorporation of cybrgreen dye using opticon 2 gri molecular biology ltd the conditions we used as recommended by manufacturer except for doubling of the concentration of the cybrgreen dye in the reaction mix 1 mkl of dynabeads was added directly to 20 mkl of pcr mix comprising primers qpcrf and qpcrx amplifying the 100 bp fragment of pask75 plasmid all 4dntps at 200 mkm each and 2 5 u of supertaq dna polymerase in 1 u00d7 taq buffer ht biotechnology we were able to reliably detect and discriminate between 10 4 and 10 9 plasmid copies plasmid elution amp transformation for plasmid elution beads were incubated in 200 u03bcl bead elution buffer beb 1 mm edta 0 1m naoh for lt 5 min beads were captured on the magnet and the plasmid in the supernatant precipitated by the addition of 1 10 v of 3m naac 3v of ethanol and mussel glycogen roche.