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Interesting salient features of our method include (a) avoidance of limiting (in treating the Hamiltonian of the level set equation), (b) anchoring the level set in a manner that ensures no drift and no spurious oscillations of the zero level during PDE-reinitialization, and (c) a non-linear tagging procedure for defining the neighborhood of the interface subject to mesh refinement.
The tagging procedure is outlined in Figure 1B.
Great care was taken during the tagging procedure to minimize detrimental effects on the fish.
For further details on tagging procedure see Skov et al. [41].
This fosmid library provides an excellent starting point for the construction of reporter genes and it is what we use for the reporter tagging procedure we present here.
During the tagging procedure, which took approximately 20 min, the gills of the fish were irrigated with water containing diluted FA 100 to maintain sedation.
Common tag SNPs (minor allele frequencies >5%) for each gene were selected using HapMap data for the Utah residents with Northern and Western European ancestry [38] and the multi-marker tagging procedure in Tagger (r2>0.8) [39].
By strictly handling the PCR tagging procedure in 96-wells format and by automating the ligation of plate-tags and library preparation, it is possible to increase the number of samples without affecting the sample handling complexity.
Advantages of the RAD-tag procedure are that no reference genome is needed and that detection of SNPs and screening their variation is performed in the same run.
DOI: http://dx.doi.org/10.7554/eLife.09579.003 The photo-tagging procedure uses two-photon microscopy (Skoch et al., 2006) rather than conventional confocal microscopy.
The total time a cricket was in captivity was 90 120min, with an additional 40min at first capture for the tagging procedure.
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