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All constructions were performed in bacteria using pBluescript as vector.
All plasmid constructions were performed by standard molecular biology techniques.
RNA isolation and library constructions were performed as previously described amplifying the VHH genes and ligate them into a phagemid vector for display on phage resulting in libraries of size around 108 [8].
Immunizations of llamas and library constructions were performed as described previously [3], [8] with six weekly injections for RSV and H5N1 and collection of 150 ml blood for isolation of RNA from the peripheral blood lymphocytes seven weeks after the initial immunization, resulting in two libraries for each viral target.
The constructions were performed by PCR-based mutagenesis, which involved two sequential steps [ 49].
All the constructions were performed in E. coli (strain XL1-Blue [ 13]) and then used to electrotransform S. oneidensis.
Similar(47)
To construct the different strains, transformation of U. maydis protoplasts with the indicated constructions was performed as described previously [66].
Secondly, all models considered will be finite, because no matter what constructions are performed, the starting point will always be a finite set of points.
Phylogenetic analysis and dendrogram construction were performed, using MEGA5 software.
Cluster analysis and dendrogram tree construction were performed based on dissimilarity matrices with the un-weighted pair-group method using arithmetic averages (UPGMA).
Alignment and tree construction were performed as described above.
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