do25142
Protocols: Nuclei was isolated from frozen livers from young C57BL/6 mice (3 month old) using the protocol described in Krishnaswami et al. 2016 (Nature Protocols, 11: 499-524). Nuclei was purified using Iodixanol gradient (Optiprep, D1556, Sigma Aldrich Chemie). Nuclei were stained with Hoechst and FACS sorted according to their ploidy level diploid (2n) and tetraploid (4n) in 384-well plates. cDNA were made on 384-well plates using single-nucleus-RNA-seq2 protocol based on a modified version of SMART-Seq v4 Ultra Low input RNA (Takara). ERCC spike-in RNA (Ambion) (0.25 µL diluted at 1:100,000 and 1:300,000) was added to the RT-Mix (5µL final reaction). Sequencing libraries were prepared using Illumina Nextera XT kit (Illumina, #FC-131-1096) and the combination of 384 Combinatorial Dual Indexes (Illumina- Set A to D , #FC-131-2001 to FC-131-2004) following the manufacturer instructions but miniaturizing the reactions 10x using a liquid handling robot (Mosquito HV).
ENA-FIRST-PUBLIC
2021-05-07T00:26:49Z
ENA-LAST-UPDATE
2020-07-20T14:59:00Z
External Id
SAMEA7073798
INSDC center name
Helmholtz Pioneer Campus
INSDC first public
2021-05-07T00:26:49Z
INSDC last update
2020-07-20T14:59:00Z
INSDC status
public
Submitter Id
E-MTAB-93331595253410:do25142
age
3
broker name
ArrayExpress
common name
house mouse
developmental stage
adult
individual
16/036335 (SNI-13)
organism part
liver
ploidy
2n
post analysis well quality
fail
sample name
E-MTAB-93331595253410:do25142
scientific_name
Mus musculus
sex
male
strain
C57Bl6/J