2.3. cDNA synthesis and qPCR of target-specific genes
Multiplexed cDNA synthesis was performed using the TaqMan1
MicroRNA Reverse Transcription kit for creating custom reverse
transcription (RT) pools (Life Technologies). For this purpose, a RT
primer pool was created consisting of individual RT primers for
miR451 and miR16 (blood-specific), miR205 and miR658 (saliva-
specific), miR10b and miR135b (semen-specific) and miR203 (skinspecific).
All TaqMan1 assays were run in triplicate.
2.4. Data analysis
LinRegPCR corrected qPCR data were normalized against the
validated references (miR92 and miR374) and against the standard
reference U6B using the software qbaseplus (Biogazelle). Finally,
normalized relative quantities (NRQs) were compared.