GSM3609706: 15: Nontargeting; Mus musculus; RNA-Seq
Lung lobes stored in RNALater solution were weighed (~25-30 mgs) and minced into 2-3 mm pieces. Total RNA was extracted with QIAzol Lysis Reagent (Qiagen, 1023537) using Zirconia beads (Biospec,11079110ZX) and a Mini-Bead Beater16™.  Total RNA, including microRNA, was isolated using a Qiagen miRNEasy kit (#217004). Three methods were used to determine RNA concentration and RNA quality -  spectrophotometry of total RNA (NanoDrop 1000), gel filtration of RNA integrity (RNA 6000 Nano Kit, Agilent 2100 Bioanalyzer), and fluorescent dye binding (Quant-iT RiboGreen Kit, Invitrogen55).  RNA quality determined by A260/280 ranged from 1.8 to 2.2, A260/230 >2 and RIN values ranged from 6.9 to 8.6. Initial RNA quantification was performed with a Qubit RNA assay kit, followed by RIN determination with an Agilent 2100 Bioanalyzer. cDNA libraries were prepared using 2 micrograms of RNA in TruSeq RNA sample preparation kits. Final quantification was performed with a Kapa Library Quant kit. Nonstranded cDNA libraries were sequenced using an Illunina HiSeq 2500 instrument (HiSeq v4 reagents) at read depth of 10M reads/sample for unsensitized and nontargeting and 15M reads for HDM and antimiR-145 treated samples.
GEO
GSM3609706
GEO Accession
GSM3609706