title: P16_B2
organism: Homo sapiens
description: Protocols: Tissues were frozen in dry-ice cooled isopentane and stored in air-tight tissue cryovials at -80°C. Prior to undertaking any spatial transcriptomics protocol, the tissues were embedded in an optimal cutting temperature compound (OCT) and tested for RNA quality with RNA integrity number (RIN). Tissues with RIN values >7 were cryosectioned in a pre-cooled cryostat at 10 μm thickness. Two consecutive sections were cryosectioned at 10 μm thickness in a pre-cooled cryostat and transferred to the four 6.5mm x 6.5mm capture areas of the gene expression slide. Slides were fixed in methanol for 30 minutes prior to staining with H&E and then imaged using the Nanozoomer slide scanner. The tissues underwent permeabilization for 24 minutes. Reverse transcription and second strand synthesis was performed on the slide with cDNA quantification using qRT-PCR using KAPA SYBR FAST-qPCR kit (KAPA Biosystems) and analysed on the QuantStudio (ThermoFisher). Following library construction, these were quantified and pooled at 2.25 nM concentration. Reverse transcription and second strand synthesis was performed on the slide with cDNA quantification using qRT-PCR using KAPA SYBR FAST-qPCR kit (KAPA Biosystems) and analysed on the QuantStudio (ThermoFisher). Following library construction, these were quantified and pooled at 2.25 nM concentration.
attributes.tag: disease
attributes.value: lung adenocarcinoma
attributes.tag: individual
attributes.value: Patient 16
attributes.tag: organism
attributes.value: Homo sapiens
attributes.tag: organism
attributes.value: Homo sapiens
attributes.tag: collection date
attributes.value: not collected
attributes.tag: disease staging
attributes.value: T2bN0M0
attributes.tag: sex
attributes.value: female
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: common name
attributes.value: human
attributes.tag: organism part
attributes.value: lung
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: sampling site
attributes.value: normal tissue adjacent to tumor
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: original source name
attributes.value: TB20.0327
attributes.tag: age
attributes.value: 90
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-11-16