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For each template, a control primer (CNV_C), which together with CNV1_F amplifies the wild-type sequence at the 3′ end of the CNV, was used in parallel to verify presence of the template.
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The PDMS microchip was created from three patterned templates: a control layer, a flow layer and a cover plate, using multilayer soft lithography.
In addition, as a template control, a 350 bp fragment of the 28S rDNA gene was amplified using the primers shown in Table 1.
Control reactions included a no template control and a control without reverse transcriptase.
Primers and probes specific for HLA-DRA were included as a template control.
All samples were tested in duplicate and each run included a template control.
Each assay run contained two negative controls (a no template control and a no reverse transcriptase control) and a standard dilution curve.
Additionally, several no template controls and a positive control in triplicate were included.
For each set of primers, a no template control and a no reverse amplification control were included.
Negative controls included a no template control and a no reverse transcriptase control.
β-Actin was amplified as a template loading control.
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