description: Protocols: PDX models were generated using subcutaneous tumor implantation in female NOD.Cg-Prkdc<scid> Il2rg<tm1Wjl>/SzJ (NSG; JAX strain 005557) host mice. Tumors were grown, collected, and fragmented for serial passage in NSG mice. Tumor samples at described passage numbers were taken for expression analysis. Tissues preserved in RNAlater were homogenized in TRIzol (ThermoFisher Scientific) using a gentleMACS dissociator (Miltenyi Biotec Inc). Total RNA was isolated using the miRNeasy Mini kit (Qiagen) according to manufacturer‚Äôs protocols, including the optional DNase digest step. RNA quality and concentration were assessed using the RNA 6000 Nano LabChip assay on the 2100 Bioanalyzer instrument and Nanodrop 2000 spectrophotometer (Thermo Scientific). Stranded libraries were prepared using the KAPA mRNA HyperPrep Kit (KAPA Biosystems), according to the manufacturer's instructions. Briefly, the protocol entails isolation of polyA containing mRNA using oligo-dT magnetic beads, RNA fragmentation, first and second strand cDNA synthesis, ligation of Illumina-specific adapters containing a unique barcode sequence for each library, and PCR amplification. Libraries were checked for quality and concentration using the DNA 1000 assay (Agilent Technologies) and quantitative PCR (KAPA Biosystems), according to the manufacturers' instructions.
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism
attributes.value: Homo sapiens
attributes.tag: age
attributes.value: 77
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: sex
attributes.value: female
attributes.tag: disease
attributes.value: cancer
attributes.tag: organism part
attributes.value: Lung
attributes.tag: individual
attributes.value: LG0645F826P0000P1015P2
attributes.tag: race
attributes.value: White
attributes.tag: ethnic group
attributes.value: Not Hispanic or Latino
attributes.tag: primary site
attributes.value: Lung
attributes.tag: specimen site
attributes.value: Lung
attributes.tag: diagnosis_initial
attributes.value: lung adenocarcinoma
attributes.tag: diagnosis_clinical
attributes.value: lung adenocarcinoma
attributes.tag: tumor type
attributes.value: Primary Malignancy
attributes.tag: tumor grade
attributes.value: NULL
attributes.tag: tumor stage
attributes.value: AJCC II
attributes.tag: passage number
attributes.value: P2
attributes.tag: treatment naive
attributes.value: Yes
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2021-12-17
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2021-12-17