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Fig. 1 | Biomarker Research

Fig. 1

From: Extracellular miR-6723-5p could serve as a biomarker of limbal epithelial stem/progenitor cell population

Fig. 1

Workflow of RNA extraction from cLEC culture media. Overview of the methodology developed to select the RNA extraction method from cLEC conditioned media. In arm 1, media is harvested every 48 hours once the average diameter of the explant outgrowth reached 7 mm. Cells and debris were removed before flash freezing. Media was concentrated (~ 22 times) using a 3 kDa exclusion column prior to RNA extraction. In arm 2, media is harvested at a unique time point corresponding to cell collection. Centrifugation allows for cell and debris removal before flash freezing. Subsequently, media is processed for RNA extraction. In both arms, total volume is divided by three and processed with three different miRNA extraction kits. RNA quality and concentration are determined using both Nanodrop and Bioanalyzer small RNA electrophoresis. A, B and C refer to mirVana PARIS, miRNeasy micro and miRNeasy Serum/Plasma Advanced kits respectively. 1 and 2 refer to the concentrated and unconcentrated arm respectively

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