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The effectiveness of the DNase digestion was assessed using controls that omitted reverse transcriptase.
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Control reactions omitting reverse transcriptase were performed to validate successful removal of contaminating genomic DNA.
"Mock cDNA" was generated by performing reactions in parallel with those used to generate cDNA, omitting reverse transcriptase enzyme from the reaction.
We also performed nRT-PCR on tissue RNA, omitting reverse transcriptase.
Negative controls of RT-PCR experiments were always performed by omitting reverse transcriptase (not shown).
Negative control samples (−RT) were prepared by omitting reverse transcriptase during cDNA synthesis.
Assays to determine DNA contamination were carried out by omitting reverse transcriptase from the reactions.
Samples from reactions omitting reverse transcriptase were used to verify that the reaction did not amplify residual DNA.
Negative controls were performed by omitting reverse transcriptase from the RT reactions to verify that the amplified products were from the mRNA and did not originate from genomic DNA contamination.
A negative control (omitting reverse-transcriptase enzyme) was included to confirm there was no genomic DNA contamination of RNA samples.
Negative controls included reverse transcription reactions that omitted the reverse transcriptase.
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