Simultaneous Detection of Different MicroRNA Types Using the ZIP-Code Array System

Table 3

miRNA-specific primers and assays used in quantitative real-time PCR. miRNA-specific commercial assays (Qiagen) and sequences of self-made miRNA-specific primers used in qRT-PCR (Tm = melting temperature).

miRBase ID	MiScript Assay (Qiagen)	Forward primer/reverse primer (5′ to 3′ orientation)	Length (nt)	Tm (°C)	Amplicon length (bp)	PCR efficiency	Error rate (%)
hsa-miR-9-5p	Hs_miR-9_1 miScript Primer Assay				85–87	2.3	4.90
hsa-mir-9-3		GAGGCCCGTTTCTCTCTTTGG/	21	59	63	1.6	3.91
		CTAGCTTTATGACGGCTCTGTGG	23	59
hsa-miR-16-5p	Hs_miR-16_2 miScript Primer Assay				85–87	1.9	2.11
hsa-mir-16-1		GCAGCACGTAAATATTGGCGTTAAG/	25	59	74	1.8	0.61
		GTCAACCTTACTTCAGCAGCACAG	24	60
hsa-miR-199b-5p	Hs_miR-199b_1 miScript Primer Assay				85–87	2.5	3.47
hsa-mir-199b	Hs_mir-199b_1_PR miScript Precursor Assay				89	1.9	0.45
hsa-miR-99a-5p	Hs_miR-99a_2 miScript Primer Assay				85–87	1.9	1.34
hsa-mir-99a	Hs_mir-99a_1_PR miScript Precursor Assay				72	1.9	4.15