GSM6697747: MCI5, Mild Cognitive Impairment, monocytes; Homo sapiens; RNA-Seq
Monocytes were first homogenized in 500µL of cold TRIzol and then 500µL of chloroform:isoamyl alcohol 24:1 (Sigma Aldrich) were added. Samples were vortexed for 20 seconds and incubated 2 min at room temperature. The samples were then centrifuged at 10,000g for 18 min at 4°C. The aqueous phase, that contains RNAs, was transferred in a new Eppendorf tube. An equal volume of ethanol 100% was added and the mixture was then loaded into a Zymo-Spin™ IIICG Column from the Quick-RNA™ Miniprep Kit. Further steps were conducted according to the manufacturer's protocol. The RNA concentration in ng/µL was determined using a Nanodrop system (Thermo Fisher Sscientific). Library preparation was initiated with 10ng of total RNA. RNA-seq libraries were prepared using NEBNext® single cell/low input RNA library preparation Kit (New England Biolabs).