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The pyrosequencing assays were prepared and performed according to the manufacturer's instructions, each assay being performed in triplicate, on three different template samples.
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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.
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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.
We prepared fifty-nine differentemplateses and measured eNOS mRNA copy numbers in Matrigel VEGF-transfectant samples.
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