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Second, and partly as a result of this new configuration of stakeholders, the long-standing debate on targeted "vertical" programs versus the "horizontal" health systems strengthening is now being played out in the aid effectiveness arena, with so-called "diagonal" approaches [ 39] guiding alignment and harmonisation of donor activities at country level [ 40].
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Sequences shorter than 100 amino acids were omitted from analyses as those tended to align poorly even in a guided alignment (data not shown).
This could affect alignment slightly, in the case of non-annotation guided alignment, for a small number of ambiguously aligning reads.
CoMFA and CoMSIA analyses were done using ligand based and receptor guided alignment schemes.
Images were deconvolved, projected and manually aligned as layers in Photoshop, using GFP filaments and follicle cell nuclei in overlapping regions to guide alignment.
First, since automated methods, like RNASalsa [ 89], remain largely underappreciated, secondary structure guided alignment still relies on extremely time-consuming manual alignment.
The insert sizes were checked by digestion with EcoRI, sequenced by Sanger sequencing, and assembled using Sequencher 4.10.1 using non-repetitive flanking sequences to guide alignment.
By providing a causal framework for proposing molecular homology, secondary structure consensus guided alignment contrasts with the methods that rely on simple sequence similarity or congruence optimization.
Reference guided alignment to both mouse and rat reference genomes and consensus sequence assembly was performed with MAQ 0.7.1 [ 22] using default settings.
Thus, quality assessment of data via e.g. secondary structure guided alignments, discarding of randomly similar aligned positions or heterogeneity of the data set prior to analysis is a crucial step to obtain reliable results.
Although the approach varies, each of these methods aligns read data to guide alignments using reference trees to determine the best homology.
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