GSM4648920: RNA-seq_hMGL TREM2-KO Rep7; Homo sapiens; RNA-Seq
After washing with cold PBS, hMGLs were lysed using TRIzol® Reagent (Thermo fisher, Cat # 15596026) and stored at -80C for RNA extraction. Total RNA was extracted using the RNeasy Plus Micro Kit (50) (QIAGEN #74034), following manufacturer's recommendations with slight adjustment. Briefly, 200 ul of chloroform was added to the TRIzol extraction solution/cells, mixed, and centrifuged. RNa was then extracted from the aqueous phase according to the manufacturer's recommendations. RNA was quantified using a NanoDrop spectrophotometer and RNA integrity numbers (RINs) were assessed using an Agilent TapeStation 4200 system; all samples used for subsequent analysis showed a minimum RIN of 9 or 10. For each replicate, RNA-seq libraries were prepared from 500 ng of purified total RNA using the NEBNext® Ultra II™ Directional RNA Library Prep Kit for Illumina®(NEB, Ipswich MA) following manufacturer's recommendations, and sequenced on an Illumina Hisdeq platform as paired end 150-bp reads (Genewiz, LLC, South Plainfield, NJ). An average sequencing depth of 45.13 M (±3.6 M) was obtained in from the hMGL samples, with minimum and maximum values of 39.2 and 64.6 M.
GEO Accession
GSM4648920