To improve the trackability of the mobile robot, we adopted a Bézier subdivision method and inserted a quintic Bézier segment into high-curvature areas.
The synthetic genes of CCR5, CCR3, CXCR4 and CX3CR1 were synthesized using a two-step assembly/amplification PCR method and inserted into two different kinds of expression systems.
For generation of transgenic lines we used the ɸC31 integrase method and inserted constructs in attP2 and attP40 docking sites (Groth et al., 2004; Markstein et al., 2008; Pfeiffer et al., 2010).
Plasmids from 97 individual clones harboring noisy yEGFP expression were extracted using a standard phenol chloroform extraction method and inserted fragments were sequenced using a primer hybridizing in the yEGFP gene 75 bp upstream of the start codon (supplementary table S7, Supplementary Material online).
For this purpose, the migration pooled according to the specific combination of prosthesis type, fixation method and articulating insert (PFI) and visualized in a dot chart (Jacoby 2006).
The whole DNA sequence was synthesized by a one-step total gene synthesis method, and then inserted in pUC18 vector.
Plasmid DNA from recombinant E. coli strains was extracted according to the TempliPhi method (GE Healthcare) and inserts were sequenced on both strands using universal forward and reverse M13 primers and the ET DYEnamic terminator kit (GE Healthcare).
With this method, the vector and insert are PCR amplified separately, with only 18 cycles, using a high fidelity DNA polymerase.
Thirdly, a speed-up evaluation method and a fast Insert-based neighborhood examining method are developed based on the properties of the no-wait FSSPs.
Plasmid DNA was isolated from the selected colonies using the alkaline-lysis method and the cDNA inserts sequenced from the 5'-end using specifically designed primer PT2F2 5'-GCGCCATTGTGTTGGTACCC-3'.
