Quantitative Reverse-Transcriptase Polymerase
Chain Reaction
Total RNApurificationwas performed on all specimens (using an
Aurum Total RNA Mini Prep-Kit; Bio-Rad, Hercules, California,
United States), including a DNase 1 digest, according to the
manufacturer’s instructions. Reverse transcriptase PCR was
performed on a portion of the purified RNA for each specimen
to create complementary DNA (cDNA) libraries using the iScript
cDNA Synthesis Kit (Bio-Rad) under the following conditions in a
conventional thermocycler: 5minutes at 25°C, 30minutes at 42°
C, and 5 minutes at 85°C.
International Archives of Otorhinolaryngology Vol. 19 No. 2/2015
Quantitative Reverse-Transcriptase PolymeraseChain ReactionTotal RNApurificationwas performed on all specimens (using anAurum Total RNA Mini Prep-Kit; Bio-Rad, Hercules, California,United States), including a DNase 1 digest, according to themanufacturer’s instructions. Reverse transcriptase PCR wasperformed on a portion of the purified RNA for each specimento create complementary DNA (cDNA) libraries using the iScriptcDNA Synthesis Kit (Bio-Rad) under the following conditions in aconventional thermocycler: 5minutes at 25°C, 30minutes at 42°C, and 5 minutes at 85°C.International Archives of Otorhinolaryngology Vol. 19 No. 2/2015
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