Mircute mirna qrt pcr detection kit sybr green

The expression patterns of the pathways involving miR169a-NF-YA1-GBSSI/SSIIIa and miR169o-GATA9-SSIIIa/SBEIIb were analyzed by qRT-PCR. Total RNAs and miRNAs from Mo17 and Ji419 at 15, 20, 25, 30, 35, and 40 DAP were used for analysis of gene expression patterns by qRT-PCR to clarify any possible regulatory relationship between the chosen miRNA-target pairs in developing maize endosperm. Two target genes, NF-YA1 and GATA9, were analyzed using qRT-PCR as described above. The first-strand cDNAs of these miRNAs were synthesized using the miRcute miRNA First-Strand cDNA Synthesis Kit (TianGen, Beijing, China). qRT-PCR was then carried out to analyze expression of these miRNAs using the Bio-Rad iQ5 (Bio-Rad, USA) following the instructions for the miRcute miRNA qRT-PCR Detection Kit (SYBR Green) (TianGen, Beijing, China). Three technical replicates were also performed, and 5S rRNA was used as internal reference. The primers used for qRT-PCR were listed in Additional file 2: Table S9.