The miRNeasy Mini Kit enables purification of total RNA which includes RNA from 18 nucleotides (nt) upwards. Alternatively, an miRNA-enriched fraction and a total RNA (>200 nt) fraction can be can be purified separately. A separate miRNA-enriched fraction may be required to reduce background in particularly sensitive downstream applications.
Total RNA was purified from 10-fold serial dilutions of Jurkat cells (106 to 10 cells) using the miRNeasy Micro Kit. RNA was purified from 2 replicates of each dilution, either [A] manually or [B] using the QIAcube. Purified RNA was used to detect miR-16 using the miScript PCR System on a Rotor-Gene Q cycler. miR-16 was detected in as few as 10 cells and CT values were consistently reproducible. No signal was detected in negative controls (blue and yellow curves).