title: D2_1
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: ENA first public
attributes.value: 2024-04-29
attributes.tag: External Id
attributes.value: SAMEA114630166
attributes.tag: INSDC center name
attributes.value: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, CB10 1SA, UK University of Cambridge, Department of Haematology, Cambridge, CB2 0AW, UK Wellcome Trust – Medical Research Council Cambridge Stem Cell Institute, Cambridge, CB2 0AW, UK
attributes.tag: INSDC last update
attributes.value: 2023-11-16T14:17:22Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-13530:D2_1
attributes.tag: age
attributes.value: 67
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: normal
attributes.tag: disease staging
attributes.value: not applicable
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: individual
attributes.value: Donor 2
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: lung
attributes.tag: original source name
attributes.value: 534C
attributes.tag: sample name
attributes.value: E-MTAB-13530:D2_1
attributes.tag: sampling site
attributes.value: healthy tissue
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male