Fig. 3

Optimization of the Direct S-Poly(T) Plus assay. a Reducing effects of hotstart Alpha Taq polymerase on the non-specific amplification, RT-qPCR products of five miRNAs (hsa-miR-93-5p, hsa-miR-15b-5p, hsa-miR-150-5p, hsa-miR-21-5p, hsa-miR-451a and hsa-miR-92a-3p) were conducted electrophoresis, -RT: no-reverse transcriptase control; b amplification plot of hsa-miR-15b-5p with normal and hot-start Alpha Taq polymerase; c amplification plot of hsa-miR-93-5p, hsa-miR-15b-5p, hsa-miR-150-5p, hsa-miR-21-5p, hsa-miR-451a and hsa-miR-92a-3p without reverse transcriptase using Alpha Taq polymerase; d amplification plot of hsa-miR-93-5p, hsa-miR-15b-5p, hsa-miR-150-5p, hsa-miR-21-5p, hsa-miR-451a and hsa-miR-92a-3p without reverse transcriptase using hot-start Alpha Taq polymerase; e correlation of different amount of hot-start Alpha Taq polymerase inputs to the threshold cycle (Ct)