Protocols: Tissue pieces were homogenized in prechilled 250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 10 mM HEPES pH 8, 1 %, BSA, 0.1 % IGEPAL, 1 µM DTT, 0.4 U/µL RNaseIn, and 0.2 U/µL Superasin and lysed for 5 min on ice. The lysate was centrifuged at 100 x g for 1 min at 4 °C. The supernatant was transferred to a new reaction tube and centrifuged at 500 x g for 5 min at 4 °C. The supernatant was removed and the pellet resuspended in 1 mg/mL DSP in PBS and incubated for 30 min at room temperature. The fixation was quenched by addition of Tris-HCl to a final concentration of 20 mM. The fixed suspension was pelleted at 500 x g for 5 min at 4 °C. The supernatant was removed and the pellet resuspended in 250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 10 mM TRIS-HCl pH 8, 1 %, BSA, 1 µM DTT, 0.4 U/µL RNaseIn, and 0.2 U/µL Superasin. This was centrifuged at 500 x g for 5 min at 4 °C. The supernatant was removed and the pellet resuspended in PBS. Nuclei were isolated through a combination of mechanical and chemical lysis. Per library 15,000 - 20,000 nuclei were loaded into one channel of the ChromiumTM Single Cell A Chip (10X Genomics®). Libraries were prepared using Chromium Single Cell 3′ Library & Gel Bead Kit v2 according to manufacturer's instructions.
ENA first public
2021-11-08
ENA last update
2021-11-08
External Id
SAMEA10418816
INSDC center alias
Center for Molecular Biology of Heidelberg University (ZMBH)
INSDC center name
Center for Molecular Biology of Heidelberg University (ZMBH)
INSDC first public
2021-11-08T12:18:22Z
INSDC last update
2021-11-08T12:18:22Z
INSDC status
public
Submitter Id
E-MTAB-11063:SN011
age
32
broker name
ArrayExpress
common name
human
developmental stage
adult
disease
normal
genotype
wild type genotype
individual
1
organism part
testis
replicate
1.2
sample name
E-MTAB-11063:SN011
sex
male