FIGURE 2.
Impact of length of target binding overhang, strand accessibility, and reaction volumes on the sensitivity of the modified protocol. The target used is the antisense strand of the GFP22 siRNA duplex. (A) Amplification using stem–loop primers with different lengths of the target binding overhang. (B) Detection of single-stranded vs. double-stranded target RNA using the stem–loop primer with a 6-nt target binding overhang. Technical triplicate, mean, and range of values (A,B). (C) Sensitivity of the method depending on the reverse transcription and PCR reaction volumes. (NTC) No template controls; (XTC) control template mature human miR146.
