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Similar to our method, they also presented an optimization problem of association control for efficiency and fairness.
To control for efficiency of transfection, 0.5 µg of pRL-TK (Promega), which expresses Renilla luciferase, was included.
The housekeeping β-actin gene was used as a control for efficiency of RNA extraction and transcription.
A Renilla reniformis luciferase plasmid (phRL-CMV-Luc from Promega) was also cotransfected as a control for efficiency.
In the proposed method, we decided to add in the mix an exogenous spike-in miRNA (cel-mir-39 coniugated with VIC fluorescence) as internal control for efficiency of the whole reaction (from extraction to PCR) since the fluorescence analysis using only FAM probes did not allow a precise quantification of miRNAs.
Similar(55)
Each PCR array contained 84 transcripts of the corresponding signaling pathway, a set of five housekeeping genes as internal controls and additional controls for efficiency of reverse transcription, PCR and the absence of contaminating genomic DNA.
Renilla was used as an internal control for transfection efficiency.
The PAT antibody was used as a control for transformation efficiency.
The reporter phRL-CMV was included to serve as a control for transfection efficiency and sample handling.
pCMV-Rluc served as a control for transfection efficiency.
The SV40-driven luciferase reporter, pGL3-Control Vector (Promega), was used to control for transfection efficiency.
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