Agilent rna 6000 nano pico chip

Both differentiated cells and controls were subjected to RNA isolation. The first step was the cell detachment with 0.25% trypsin solution. Subsequently, cells were suspended in 1 mL of TRIzol (Thermo-Fischer Scientific, Waltham, MA, USA) and immediately frozen at −80 °C. RNA was isolated using phase separation with chloroform, and isopropanol was used for RNA precipitation from the aqueous phase. Then RNeasy Mini kit was utilized for total RNA purification. RNA was eluted in 30 µL of RNAse/Dnase free water, subjected to quality assessment and stored at −80 °C. The Qubit™ RNA BR/HS Assay Kit (Thermo Fisher Scientific, Waltham, MA, USA) and the Agilent RNA 6000 Nano/Pico Chip (Agilent, Santa Clara, CA, USA) and the Bioanalyzer 2100 instrument (Agilent, Santa Clara, CA, USA) were used to RNA quantification and quality assessment. Both RIN values (6.9–10) and the concentration (6.2–335.0 ng/µL) fulfilled the criteria for library preparation.

After differentiation, cells destined for RNA isolation (both the differentiated cells and controls) were detached using a 0.25% trypsin solution and suspended in 1 mL of TRIzol (Thermo-Fischer Scientific, Waltham, MA, USA) and immediately frozen at −80 °C. After phase separation using chloroform, total RNA was precipitated from the aqueous phase by adding isopropanol. Then, the total RNA was purified using an RNeasy Mini kit, eluted in 30 µL of RNAse/DNase free water, and stored at −80 °C after quality assessment. Quantification of the isolated RNA and its quality was performed using the Qubit™ RNA BR/HS Assay Kit and the Agilent RNA 6000 Nano/Pico Chip on the Bioanalyzer 2100 instrument, respectively. Both the concentration (6.2–335.0 ng/µL) and RIN values (6.9–10) met the criteria for library preparation.