strain: SCL-tTA H2B-GFP transgenic mouse line that expresses the fusion protein histone H2B green fluorescent protein (H2B-GFP) under the control of a tetracycline responsive regulatory element, with the expression of the tTA-S2 transactivator under the control of the endogenous scl locus (SCL-tTA) limiting expression of GFP to both early embryonic and adult HSC/progenitor cells. tissue: Bone Marrow sorted for non-LRC CD34 Neg, Lin Neg, Sca1+, cKit+, CD150+, CD48 Neg, GFP Neg HSCs timepoint: After 170 day chase (+ doxycycline)
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated from bone marrow sorted for non-LRC CD34 Neg, Lin Neg, Sca1+, cKit+, CD150+, CD48 Neg, GFP Neg HSCs, using the PicoPure RNA kit (Arcturus, Menlo Park, CA). Per condition 3 independent samples were analysed. Two rounds of amplification for each RNA sample were performed utilizing the Nugen WT-Ovation Pico RNA Amplification System (Nugen, San Carlos, CA, USA).
Label
Biotin label
Label protocol
Biotin labelling of cRNA was performed utilizing the Affymetrix GeneChip IVT labelling kit (Affymetrix, Santa Clara, CA, USA).
Hybridization protocol
Hybridization performed on a Affymetrix Fluidics Station
Scan protocol
Array scanning and raw data processing were done using GCOS software (Affymetrix)
Description
Murine non-LRC HSC Replicate 3
Data processing
Data processing was performed utilizing the DNA Array Facility of Lausanne’s Remote Analysis System (http://race.unil.ch), a web-based interface for various statistical analysis routines performed utilizing the R language (http://www.r-project.org). Data was normalized using RMA.
