Quantitative RT-PCR Specific for Precursor and Mature miRNAs - miRNA Maturation: Methods and Protocols - page 121

Fig. 3 Total RNA quality analysis via BioAnalyzer. Shown are representative electropherograms and gel-like
images from tissue preparations using the RNA 6000 Pico LabChip kit and the 2100 BioAnalyzer. RIN RNA
integrity value

1. Adjust RNA to a fi nal concentration of 5 ng/ μ L and place on

3.4 cDNA Synthesis
for Mature miRNA ice ( see Notes 7 and 8 ).

2. Prepare a reverse transcription master mix (RT master mix) for
the selected strategy ( see Note 9 ):
TaqMan Reverse Transcription Kit components for stem-loop
qRT-PCRs ( see Table 1 ).
Qiagen miScript Reverse Transcription Kit for polyT adaptor
qRT-PCRs ( see Table 2 ).