title: 5C of p0wt_liver
organism: Mus musculus
description: Single cells were crosslinked with 1% formaldehyde for 10 min at room temperature. The crosslinked DNA was lysed and digested with HindIII. The DNAs were then ligated with T4 DNA ligase and purified with phenol-chloroform. 500 ng of the ligated DNA was mixed with 1 μg of salmon testis DNA (Sigma). 1.7 fmol of each 5C primer and 1 μl of 10×5C annealing buffer (NEBuffer 4) were added into the samples to make a total volume of 10 μl. Denaturing was performed by incubating samples at 95 °C for 5 min followed by annealing at 48 °C for 16 h. The DNA was then ligated by Taq DNA ligase (NEB) in the Taq DNA Ligase Reaction Buffer (NEB) at 48 °C for 1 h followed by incubation at 65 °C for 10 min to stop the reaction. 6 μl of ligated products was amplified by PCR using KOD-Plus-Neo (TOYOBO) with specific primers in 25 ul of volume and purified with MinElute PCR Purification Kit (QIAGEN).
attributes.tag: strain
attributes.value: C57BL/6
attributes.tag: isolate
attributes.value: The liver of p0 mouse was directly resuspended in DMEM supplemented with 10% FBS by pipetting. Cells were then filtered through a 100um cell strainer (BD) to obtain single-cell suspension.
attributes.tag: age
attributes.value: p0
attributes.tag: sex
attributes.value: not determined
attributes.tag: tissue
attributes.value: liver
attributes.tag: biomaterial_provider
attributes.value: Qiang Wu, Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, No.800, Dong Chuan Rd. Shanghai, 200240
attributes.tag: collected_by
attributes.value: Yuanxiao Tang
attributes.tag: collection_date
attributes.value: 2015-10-18
attributes.tag: genotype
attributes.value: wild type
attributes.tag: geo_loc_name
attributes.value: China:Shanghai
attributes.tag: isolation_source
attributes.value: not collected
attributes.tag: BioSampleModel
attributes.value: Invertebrate