P-EMBL-171 - P-EMBL-171
Cells isolated from 4 independent donors for each culture condition (BM-MSC-M1, BM-MSC-M2, AT-MSC-M1, CB-MSC-M3) were harvested upon reaching 70% confluency. In analogy RNA of HS68 cells was isolated four times. About 2x10^6 cells from each fraction were lysed and total RNA isolated using the Rneasy Mini Kit (Qiagen, Hilden, Germany). DNase treatment was performed (Quiagen). RNA quality was controlled with the RNA 6000 Pico LabChip kit (Agilent, Waldbronn, Germany). Linear amplification of 10 ug total RNA was performed by a one round in vitro transcription using the Arcturus RiboAmp Kit (Acturus, Mountain View, USA). RNA of HS68 cells was always amplified next to the MSC samples. The amplified RNA was analyzed by the RNA Nano Lab Chip kit (Agilent) and by the SpectraMAX plus photometer (Molecular devices, Sunnyvale, USA) at 260 nm. We gained about 100 ug aRNA with a continuous spectrum of RNA length of 200 to 1200 base pairs.
Amplification, Extracted product