Skip to main content
. 2015 Mar 16;43(12):e77. doi: 10.1093/nar/gkv218

Figure 4.

Figure 4.

Db-PCR to specifically quantify miR-16. (A) Sequences and secondary structures of the 5′-Dbs-adapter (orange), 3′-Db-adapter (green) and targeted human miR-16 (black). The 5′-Dbs-adapter contains two 3′-terminal RNA nucleotides and a spacer in the loop region. Three different 5′-Dbs-adapters containing 6, 12 or 16 nucleotides protruding from the 5′-end were used. The regions from which primers and TaqMan probe were derived are also shown. (B) Db-PCR using a 5′-Dbs-adapter with 6, 12 or 16 nucleotides protruding from the 5′-end was applied for the detection of synthetic miR-16. The 5′-Dbs-adapter with 16-nt protruding end showed the highest detection efficiency. The data represents the average Ct value from three independent experiments with bars showing the SD. (C and D) Db-PCR was applied for the detection of synthetic miR-16 and its 5′- and 3′-variants whose sequences are shown above the graph. miR-16 was the only RNA species amplified by the Db-PCR. The data represents the average Ct value from three independent experiments with bars showing the SD. Northern blot detection of the quantified RNAs is shown below. (E) Proportional correlation of HeLa total RNA input (0.313, 0.625, 1.25, 2.5 and 5 ng) to the Ct value obtained by Db-PCR. Each data set represents the average of three independent experiments with bars showing the SD.