Liver_Input_Ago2_BRF1_2
Mus musculus
Protocols: Liver was isolated from C57BL/6J wild-type mice at 8 weeks of age Mice were maintained on a 12 hr light / dark cycle in a temperature-controlled (22ºC) barrier facility with free access to water and a normal diet (NCD, CA-1, CLEA Japan, Inc.). Liver was solubilized in TNE buffer for 30 min at 4ºC. 150 mg of protein lysate was incubated with 180 µg of antibodies against Cnot3 (Takahashi et al., 2015), Ago2 (018-22021; Wako), and BRF1 (Cell Signaling) for 1 hr at 4ºC, and then incubated with 1.2 mL of Dynabeads (Invitrogen) for 2 hr at 4ºC. mRNAs in immune complexes were isolated using Isogen II (Nippongene). 100 ng of total RNA was used for RNA-seq library preparation with TruSeq Stranded mRNA Library Prep Kit for NeoPrep (NP-202-1001; Illumina), according to the manufacturer's protocol with minor modification and optimization as follows. Custom dual index adaptors were ligated at the 5' and 3'-end of library, and PCR was performed 11 cycles.
ENA-FIRST-PUBLIC
2018-10-30T17:01:51Z
ENA-LAST-UPDATE
2018-06-22T11:47:53Z
External Id
SAMEA4753534
INSDC center name
Cell Signal Unit, Okinawa Institute of Science and Technology Graduate University
INSDC first public
2018-10-30T17:01:51Z
INSDC last update
2018-06-22T11:47:53Z
INSDC status
public
Submitter Id
E-MTAB-6941:Liver_Input_Ago2_BRF1_2
age
8
broker name
ArrayExpress
common name
house mouse
diet
normal diet
disease
normal
genotype
wild type genotype
organism part
liver
phenotype
wild type phenotype
sample name
E-MTAB-6941:Liver_Input_Ago2_BRF1_2
scientific_name
Mus musculus
sex
male
strain
C57BL/6J