By modifying the procedure for in utero electroporation of rodents, we recently created a procedure for in utero electroporation to express transgenes in post-mitotic neurons of the ferret cerebral cortex (Kawasaki et al., 2012).
We modified the procedure of John et al. (John, J., Sohmen, R., Feuerstein, J., Linke, R., Wittinghofer, A., and Goody, R. (1990) Biochemistry 29, 6058-6065) for the preparation of nucleotide-free ras such that the procedure can now be applied to 1000-fold smaller quantities of protein.
To this aim, we have slightly modified the procedure of section 4.3 in such a way that, at each iteration, only one keypoint of each match at a time is manipulated.
Because of length variations between the sequences, we modified the procedure of Ding et al. [ 48] and defined two sequences as paralogs if the aligned regions were over 70% of the shorter sequences.
To observe growth kinetics within the host, we modified the procedures of Uchiyama et al.[ 49].
In order to make the dual fluorescence assay more convenient, we modified the procedures of plasmid construction so as to speed up the workflow.
Synthesis began with acetylation of the phenol moieties of fluorescein, modifying the procedure reported by Tour using acetic anhydride and pyridine (>15 g scale, >95% yield), with a mild acid wash being the only work-up necessary (Scheme 1).
We modified the procedure by determining the number of PCs based on when the proportion of variance in the selected genes explained by the PCs exceeds 95%.
To avoid this direct exposure of FePt film to Ar plasma, we modified the fabrication procedure of patterned media as illustrated in Figure 1b.
