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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.
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Control reactions included a no template control and a control without reverse transcriptase.
In addition, as a template control, a 350 bp fragment of the 28S rDNA gene was amplified using the primers shown in Table 1.
Each analysis included a control without template DNA.
A control without template was included in all batches.
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.
For each gene a no-template control and a no-amplification control (0.01% SDS added to the reaction mixture) was included in each run.
Each assay (in triplicate) included a non-template control and a non-RT control.
Each PCR run included a no-template control and a sample without reverse transcriptase.
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