MANUAL VS MACHINERY SMALL RNA EXTRACTION BY USING A QIACUBE® MACHINE : Two methods. Two volumes.

Abstract: Sepsis is a serious condition caused by a dysregulated immune response of the host triggered by an infection that can potentially lead to malfunction of various organs or even death in severe cases. Some studies have shown that the use of biomarkers could aid in early diagnosis as well as early treatment of sepsis patients. Furthermore, various studies have investigated the idea of using extracellular microRNAs as biomarkers for sepsis diagnosis. This study aimed to see if there were any differences in the quantity and purity of small RNA -which includes microRNA- by performing two different RNA extraction methods (manual and machinery by using a QIAcube) as well as two different volumes by using the ExoRNeasy Serum/Plasma Midi Kit. Blood samples were collected solely from the same self-assessed healthy donor. The plasma samples were frozen and then thawed before the RNA extraction, whether manually or machinery by the QIAcube. The extracted small RNA was then measured for quantity and purity. The quantitative results were analysed by ANOVA followed by post-hoc Tukey test to show the statistically significant difference in the concentration of small RNA. The QIAcube showed higher concentration values compared to the manual method as well as larger initial plasma volume in comparison to the lower initial plasma volume. Meanwhile, the Kruskal-Wallis test showed no statistically significant difference in the purity values among the different methods and volumes. In conclusion, based on this study, the QIAcube could do what human hands do.