GSM4711262: PDAC_Cold_epithelium_3; Homo sapiens; RNA-Seq
FFPE blocks containing human primary tumors were serially sectioned onto Superfrost Plus microscope slides and glass slides with PEN membrane (Leica, catalog 11505158). Sections on Superfrost Plus microscope slides were used for automated immunohistochemistry to identify tumor epithelium and other cell types. PEN membrane slides were air dried and stored at -80 °C until analysis. These slides were thawed at room temperature prior to use and deparaffinized in fresh xylene for 30 seconds twice, followed by rehydration in ethanol (30 seconds each in 100%, 100%, 95%, and 70% ethanol). Slides were then transferred to RNase-free water and stained with Mayer's hematoxylin for 10 seconds. Slides were washed with RNase-free water and dehydrated by submerging them in 95% ethanol, 100% ethanol, and xylene, twice in each solution for 30 seconds. Specific regions of dehydrated slides were then microdissected by laser capture using a LMD7000 microscope (Leica). RNA from cold and hot tumor stroma and epithelium was collected using a NucleoSpin total RNA FFPE XS kit (Takara, catalog 740969.50). Sequencing libraries were prepared using a SMARTer stranded total RNA-Seq kit v2 (Takara) following manufacturer's protocol and analyzed on a NextSeq 500 sequencing system (Illumina).
GEO Accession
GSM4711262