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The cut-off value of 83% was selected to define distinct operational protein families (OPF) based on the analysis of the phylogenetic trees of total clones in combination with Mothur analysis.
Finally, the three entry clones, in combination with the modified destination vector, were incubated to create a BHD-pDESTR4R3 targeting construct through BP recombination reaction.
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An assay of the recombinants using sub-clones in combination with sequence analysis delimited the Bph15 gene to a genomic segment of approximately 47 kb on chromosome 4.
The robustness of the yeast recombination cloning in combination with high efficiency gene targeting system allows high-throughput functional genomics studies in Z. tritici.
The same construct was cloned in combination with cre recombinase to express cre recombinase in hypertrophic chondrocytes as a useful tool to study hypertrophy and endochondral ossification in vivo[ 36, 37].
Again, ten clones were sequenced with all clones carrying in combination with the round 1 mutations E104K and G238S, an extra mutation, M182T (T->C transition) with no silent or other mutations.
The use of an antibody conjugated with a fluorescent molecule, which is the target to the membrane-retained therapeutic proteins has been used to find high-producing clones in the combination with FACS.
An RT-PCR based cloning approach in combination with RACE was initially used to define the range of CYP4 genes expressed in C. gibbosum.
The FISH analysis using biotinylated DNA of RP11-190J8 BAclonene (AC021600) in combination with the digoxigenin-labelled centromeric probe for chromosome 12 allowed the characterisation of the breakpoint.
Recently, effector genomics is emerging as an efficient tool to accelerate R gene cloning, often in combination with small-scale genetic mapping and allele-mining [ 14].
To our knowledge, this is the first report utilizing the robustness of the yeast recombination cloning approach in combination with efficient targeted integration in the genome of Z. tritici.
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