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In order to assess sensitivity, reproducibility as well as limitations of the barcode tiling approach, we prepared four different template pools with engineered concentrations of individual pGIPZ shRNA expression plasmids.
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Three dimensional structure of AHAS protein in the WT was modeled based on four different templates, 3E9Y A (Arabidopsis thaliana ALS in complex with Monosulfuron), 1YBH A (A. thaliana ALS in complex with Sulfonylurea), 1JSC A (Yeast ALS as a target for herbicidal inhibitors), 1NOH A (Yeast ALS in complex with Sulfonylurea).
In preliminary experiments we determined the number of cycles that allowed the linear simultaneous amplification of the four different templates (up to 40 cycles).
In this paper, two different template modes, the rectangular model and circular model, were used according to the tracked object's template.
As described in Section 4.4.1, we identified five different template structures for SPL use cases: Tags, Specific Section, Alternative Scenarios, Step Identifier and Advice use case.
Two different template switching oligos were employed for each developmental stage.
Primer efficiency was estimated from a standard curve using five different template concentrations (12.5-200 ng); all analyses were run simultaneously in triplicates.
The pyrosequencing assays were prepared and performed according to the manufacturer's instructions, each assay being performed in triplicate, on three different template samples.
We prepared fifty-nine differentemplateses and measured eNOS mRNA copy numbers in Matrigel VEGF-transfectant samples.
Since no previous information was available for ggTas2r1 binding site and its agonist-bound conformation, two ggTas2r1 models were generated using two different templates (β2AR and M2R).
Three mesoporous carbon spheres (MPCs) are synthesized using three different templates with sizes of 22, 35 and 50 nm, respectively.
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