GSM1163072: HD291 line after 15 enzymatic passages; Homo sapiens; RNA-Seq
In mechanically passaged culture, the hESC colonies were exscinded carefully using a scalpel. In enzymatic-passaged culture, the singles cells were released as described above. As SSEA4 is one of most obstinate pluripotence markers (Ramirez et al., 2011), hESC cells were purified and enriched using the magnetic beads Dynabeads® SSEA-4 (Invitrogen) and the manufacture’s instructions were rigorously followed. The quality control of the purification showed high efficiency and no over-selection of SSEA4+ sub-population.  Total RNA was purified from cell culture using the RNeasy Mini Kit (Qiagen, Courtaboeuf, France) following manufacture’s instructions. We added an RNAse-Free DNase step in RNA extraction and an RNsae A treatment in DNA extraction to eliminate residual DNA or RNA. QC of the quantity of the total RNA using nanodrop. Library construction using TruSeq RNA Sample Prep Kits. Library Quality Control and quantification by Bioanalyzer Agilent 2100, qPCR and Picogreen.
GEO Accession
GSM1163072