title: GSM3465874: LungMF E18.5 WT-3; Mus musculus; RNA-Seq
library_construction_protocol: The CD45+F4/80loCD11blo lung preAMs were sorted by FACS Aria II flow cytometer. Total RNA was extracted from the sorted cells with miRCURY RNA Isolation Kit - Cell & Plant (Exiqon). cDNA library was generated using SMART-Seq v4 Ultra Low Input RNA Kit (Clontech). cDNA was pre-amplified by a thermal cycler (Applied Biosystem) using 12-14 cycles and sheered into 150-400 bp fragments using Bioruptor Pico sonicator (Diagenode). The DNA ends were repaired using End-It™ DNA End-Repair Kit (Epicentre). The end repaired DNA fragments were ligated to adaptors by T4 ligase (NEB) after a single “A” was added at 3' by Klenow Fragment (3'→5' Exo-) (NEB). MinElute Reaction Cleanup Kit (Qiagen) was used for DNA cleanup after each step. Illumina P5 and P7 primers with individual adaptors were used for final library amplification. The 150-400 bp final product was purified on a 2% E-gel (Thermo Fisher Scientific).
attributes.tag: GEO Accession
attributes.value: GSM3465874