The miRCURY LNA miRNA QC PCR Panel contains 6 LNA-enhanced primer sets for quality control of RNA samples (light gray) and PCR efficiency (dark gray). In addition, 5 LNA PCR primer sets that target miRNAs are included. These assays target: miRNAs expressed in most tissues (blue), miRNAs expressed in kidney (purple) and brain (green). Primer sets targeting blood miRNAs are also part of the panel (red). These assays can be used as controls of hemolysis.
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
Workflow and decision tree for determining the quality of RNA samples.