mmus-UN_1-3-skin-b2
Mus musculus
Protocols: The dorsal skin from untreated, TPA-treated, and DMBA/TPA-treated mice was dissected, and a spatial grid of 1 mm full-thickness punch biopsies was collected. The grid spanned approximately 2 by 2 cm of dorsal skin. Biopsies for nucleic acid isolation were placed into deep-well 96-well plates on dry ice immediately after collection. DNA and RNA were co-extracted from skin biopsies and liver samples using a modified protocol combining the AllPrep DNA/RNA Micro Kit (Qiagen) and the DNeasy Blood and Tissue Kit (Qiagen). Samples were homogenized in 350 μl Buffer RLT supplemented with 1% β-mercaptoethanol using the TissueLyser II (Qiagen) at 30 Hz for 2 × 3 min. Homogenates were briefly centrifuged, and 700 μl proteinase K mix (12 μl proteinase K in 688 μl nuclease-free water) was added. Lysates were incubated at 56°C for 10 min, resulting in a final volume of 1038 μl. Each sample was then divided equally for RNA and DNA extraction (519 μl per fraction). To isolate total RNA, half of each lysate (519 μl) was mixed with 260 μl absolute ethanol and applied to RNeasy MinElute spin columns (Qiagen). Columns were washed once with Buffer RW1, followed by on-column DNase digestion for 15 min at room temperature. Subsequent washes were performed with Buffer RW1, Buffer RPE, and 80% ethanol. Residual wash solution was removed by centrifugation at maximum speed for 5 min. RNA was eluted in 14 μl RNase-free water and stored at −80°C. RNA quantity and integrity were evaluated using the Agilent TapeStation High Sensitivity RNA ScreenTape assay (Agilent Technologies). To isolate total RNA, half of each lysate (519 μl) was mixed with 260 μl absolute ethanol and applied to RNeasy MinElute spin columns (Qiagen). Columns were washed once with Buffer RW1, followed by on-column DNase digestion for 15 min at room temperature. Subsequent washes were performed with Buffer RW1, Buffer RPE, and 80% ethanol. Residual wash solution was removed by centrifugation at maximum speed for 5 min. RNA was eluted in 14 μl RNase-free water and stored at −80°C. RNA quantity and integrity were evaluated using the Agilent TapeStation High Sensitivity RNA ScreenTape assay (Agilent Technologies).
ENA first public
2026-06-12
INSDC center name
German Cancer Research Center (DKFZ)
INSDC status
public
Submitter Id
E-MTAB-15743:mmus-UN_1-3-skin-b2
age
18
broker name
ArrayExpress
collection date
not collected
common name
house mouse
developmental stage
adult
genotype
wild type genotype
geographic location (country and/or sea)
not collected
individual
1
isolate
not applicable
organism part
back skin
sample name
E-MTAB-15743:mmus-UN_1-3-skin-b2
scientific_name
Mus musculus
sex
female
strain
C57BL/6J