In terms of numerical stability, more efficient stability restrictions were obtained as in the case of multi-dimensional explicit schemes.
As in almost all optimization problems with conflicting objective functions, in this study, different optimal solutions (efficient designs) that meet required restrictions are obtained.
In the first case, no parameter restrictions were assumed while in the second case, parameter restrictions were obtained by assuming limiting values for the slope of the self-thinning lines based on the average square spacing S = 100 / N (Garcia, 2009).
The double stranded (CTG 16 sequence flanked by SpeI and BglII restriction sites was obtained by hybridizing the 5'ctagt ctg)16andnd 5'gatct(cag)16a oligonucleotides, called (CTG 16 and anti- CTG 16.
The MaPIP1 1 ORF, including engineered NcoI/ SpeI restriction sites, was obtained using gene-specific primers.
All primers were obtained from Sigma-Genosys (Haverhill, Suffolk, UK), and restriction enzyme was obtained from New England Biolabs (Hitchin, Hertfordshire, UK).
A PCR product of G. stearothermophilus T6 xylanase with a His-tag and BspHI (5' terminus) and KpnI (3' terminus) restriction sites was obtained [ 40, 47, 48, 67] and inserted into the pET9d vector.
A random sub-sample of all restriction fragments was obtained through a pre-selective amplification using primers E-t and M-c, followed by four selective amplifications using primer pairs E-tct/M-cga, E-tcta/M-ctt, E-tag/M-cag, and E-tgc/M-cat (Table 3), with each E primer fluorescently labeled with either VIC or 6FAM fluorescent dyes.
PCR products were obtained as follows: PCR1 fragment (351 base pairs (bp)), comprising in the following order part of the bceA promoter, the bceA start codon followed by 6 histidine encoding codons, a PmlI restriction site (CACGTG, where CAC is the last histidine codon) and an AcsI restriction site (GGCGCGCC), was obtained using Pbcea1 and Pbcea2-ml primers.
The DNA fragment encoding transactivation domain of p53 (residue 15 29) and two restriction sites (underlined) was obtained by thermal cycling using following oligonucleotides: p53NcoBack (5′-gg aat t CC ATG GCT AGT CAG GAA ACA TTT TCA GAC CTA TGG AAA C-3′) and p53NotFor (5′- g gga ttc t GC GGC CGC GTT TTC AGG AAG TAG TTT CCA TAG GTC TG-3′).
