Protocols: CD34+ bone marrow cells (StemCell Technology) were thawed and prestimulated in culture for 24h. Cells were transduced with lentivirus of shGPR and shRenilla. Positively transduced cells were marked by GFP and sorted using the FACS Aria III (Becton Dickinson) at the MPI FACS facility, Freiburg, Germany. The following sort parameters were used: 100 um nozzle with approximately10,000 evt/s. 1000 GFP+ cells were collected into ice-cold RNA lysis buffer (ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen)) and stored at -80C until further usage. Total RNA isolation was performed from the indicated populations using ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen) according to the manufacturers instructions. DNAse treatment was performed using RNAse-free DNAse Set (Qiagen). Total RNA was used as starting material for cDNA-libraries preparation using the Low Input RNA: cDNA Synthesis, Amplification and Library Generation (NEB #E6420). 14 PCR cycles were used for cDNA amplification and 10 cycles for PCR enrichment.
ENA first public
2022-01-01
ENA last update
2022-01-01
External Id
SAMEA7699206
INSDC center alias
Max Planck Institute of Immunobiology and Epigenetics (MPI-IE)
INSDC center name
Max Planck Institute of Immunobiology and Epigenetics (MPI-IE)
INSDC first public
2022-01-01T00:12:38Z
INSDC last update
2022-01-01T00:12:38Z
INSDC status
public
Submitter Id
E-MTAB-9863:SH_Knockdown_2
age
20 to 30
broker name
ArrayExpress
cell type
hematopoietic stem cell
common name
human
developmental stage
adult
disease
normal
genotype
GPRC5C -/-
individual
5
organism part
bone marrow
sample name
E-MTAB-9863:SH_Knockdown_2
sex
male
specimen with known storage state
Frozen -80C