Taqman microrna assays specific for selected mirs

Bulge-LoopTM miRNA qPCR Primer Sets (RiboBio) were used to detect selected miRs expressions by qRT-PCRs with iTaq TM Universal SYBR Green Supermix (BIO-RAD) as described elsewhere [8 (link)]. Reverse transcription of the miRs into cDNA was done with the TaqMan microRNA reverse transcription kit (Thermo Fisher, Dubai, UAE) and TaqMan microRNA assays specific for selected miRs (Applied Biosystems, Thermo Fisher, Dubai, UAE) according to the manufacturer’s recommendations. Owing to several PCR sessions to analyze a high number of samples, we created a reference sample by pooling a fraction of all control and CHF samples. This reference sample was run in each PCR session to minimize the technical variability in our samples. All qRT-PCR reactions were performed in triplicate, and the signal was collected at the end of every cycle. All miRs expressions were calibrated against spike-in cel-miR-39, which lacks sequence homology to human miRs.

Bulge-Loop TM miRNA qPCR Primer Sets (RiboBio) were used to detect selected miRs expressions by qRT-PCRs with iTaqTM Universal SYBR Green Supermix (BIO-RAD) as described elsewhere [28 (link)]. Reverse transcription of the miRs into cDNA was achieved with the TaqMan microRNA reverse transcription kit (Thermo Fisher, Dubai, UAE) [29 (link)] and TaqMan microRNA assays specific for selected miRs (Applied Biosystems, Thermo Fisher, Dubai, UAE) according to the manufacturer’s recommendations. Owing to several PCR sessions to analyze high number of samples, we created a reference sample by pooling a fraction of all control and CHF samples. This reference sample was run in each PCR session to minimize the technical variability in our samples. All qRT-PCR reactions were performed in triplicate, and the signal was collected at the end of every cycle. All miRs expressions were calibrated against spike-in cel-miR-39, which lacks sequence homology to human miRs.