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Negative PCR controls (no cDNA template) were prepared to detect possible contamination.
Negative PCR controls (no cDNA template) were prepared, in order to detect possible contamination.
Reactions not including the SuperScript II reverse transcriptase, or cDNA template, were prepared as negative controls and analyzed in parallel.
Dilutions of the cDNA template were prepared with each tissue and amplified in triplicate using SensiMix Plus SYBR + FLUORESCEIN (Quantace Ltd., London, UK).
To reduce variability among replicates, PCR premixes, which contain all reagents except for the template, were prepared and aliquotted into 0.2 ml thin well plates.
To test the limit of detection (LOD) and quantification (LOQ) of the established quadruplex real-time PCR method, four gradient dilutions of DNA template were prepared.
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For that purpose a new hierarchical template is prepared.
Next, linear DNA template was prepared using P22 and P23 to be used as a template for in vitro transcription.
This template was prepared by a new raw material: silica gel chromatography plate (aluminum foil).
The porous template was prepared from carbamide granules and then impregnated with epoxy resin.
AAO template was prepared by a two-step anodization method.
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