mRNA-seq, 00h RA-induction KH2 cells, replicate 1
Mus musculus
mRNA-seq, 00h RA-induction KH2 cells, replicate 1
strain
KH2 ES Cells, Cat. No. MES4304
age
Uninduced
dev_stage
Embryonic
sex
male
tissue
ES Cells
biomaterial_provider
Open Biosystems, Huntsville, AL, USA
cell_line
KH2
cell_subtype
wild-type
growth_protocol
KH2 ES cells with epitope tagged Hoxa1 (3XFLAG-MYC) were grown in feeder free condition using N2B27+2i media supplemented with 2000 U/ml of ESGRO (Millipore). N2B27+2i media consist of neurobasal media (21103-049, Invitrogen) , DMEM/F12 media (10565-018, Invitrogen), 0.5X N2 (17502-048, Invitrogen), 1X B27 (17504044, Invitrogen),1X ?-mercaptoethanol (ES-007-E , Millipore), 1X Glutamax (10378-016) ,1X NEAA (07600, SCT), 3µM CHIR99021 (4423, Tocris), PD0325901 (72184, SCT), 0.033% BSA (15260037) Thermo Fisher scientific) (Qi-Long, 2008). Cells were seeded on gelatinized plate without a feeder layer. Uninduced ES cells were grown in N2B27+2i up to 80-90% confluency.
sample_type
cell culture
label
mRNA-seq, 00h RA-induction KH2 cells, replicate 1
treatment
RNA was extracted using Directzol ( Zymo Research , R2070) as per manufacturer’s instruction. Total RNA was quantified in nanodrop. Quality of RAN were assessed using bioanalyzer. RNA greater than 9.5RIN( RNA integrity Number) were used for library preparation. Poly-A selected directional TruSeq libraries were prepared according to manufacturer''s specifications.
biological_replicate
1
BioSampleModel
Model organism or animal