Control of 5C
Mus musculus
5 ng of BAC library was mixed with 1.5 μg of salmon testis DNA. .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 (Table S1) in 25 ul of volume and purified with MinElute PCR Purification Kit (QIAGEN).
isolate
Bacterial artificial chromosomes (BACs) covering the Pcdh clusters were also digested with HindIII as a control. The DNAs were then ligated with T4 DNA ligase and purified with phenol-chloroform.
sex
not determined
tissue
not applicable
biomaterial_provider
Qiang Wu, Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, No.800, Dong Chuan Rd. Shanghai, 200240
collected_by
Ya Guo
collection_date
2014-11-08
geo_loc_name
China:Shanghai
isolation_source
not collected
age
missing
dev_stage
missing
BioSampleModel
Model organism or animal