The miRNAs on the panel are expressed in many sample types. In this example, human serum, plasma, urine, colorectal cancer (FFPE) and mouse brain (fresh frozen) samples were analyzed.
miRCURY LNA miRNA QC PCR Panels offer a convenient way of determining RNA yields. In this example, a plasma sample was purified 5 times using three different RNA isolation methods. RNA spike-ins from the RNA Spike-in Kit were added to the lysis buffers of the isolation kits according to the kit handbook. RNA isolation B gives the highest yield, as indicated by the lower Cq values. Overall PCR efficiency is similar between the methods, as determined by the similar UniSp3 results. The high Cq value of UniSp6 from isolation A could indicate the presence of a PCR inhibitor in this sample.
The data from the red blood cell-specific miR-451 and the stable miR-23a can be used to monitor hemolysis. Shown is the difference in the level (ΔCq) of miR-23a and miR-451 for 34 plasma samples. A ΔCq(miR-23a – miR-451) lower than 5 in human serum or plasma represents non-hemolyzed samples. If the ΔCq is close to or higher than 7, there is an increased risk that the samples are affected by hemolysis. In case of high levels of hemolysis, miRNAs from red blood cells will make a significant contribution to the overall miRNA profile identified. Special care must be taken in the analysis of such samples
The RNA Spike-In Kit, for RT was used for qPCR quality control of commercially available plasma from two different sources (n=2 for each). The UniSp2 / UniSp4 / UniSP5 RNA Spike-in mix was introduced in the RNA isolation step (blue), and the UniSp6 / cel-miR-39-3p RNA Spike-in mix was added to the RT reaction (red), as described in the kit handbook. As seen from the Cq values, the kit offers a wide dynamic range, which is ideal for control of both the RNA isolation and RT reaction steps Moreover, the results show that the RNA isolation and the RT reaction were highly reproducible.