3'-end targeted sequencing of 6 control and 6 TDP-43 KD samples in HEK293

ABSTRACT: HEK293 FlpIn T-Rex cells were maintained in Dulbeccos Modified Eagle Medium (DMEM) with 10% fetal bovine serum (FBS), supplemented with 3 g/ml blasticidine and 50 g/ml zeocin. For the siRNA-induced knockdown of TDP-43, 20 nM of TDP-43 stealth siRNA was mixed with 10 L of RNAiMAX following the manufacturers reverse transfection protocol and added to a 10 cm dish of HEK FlpIn cells. For the non-targeting siRNA also 20 nM was used, to distinguish off-target effects from biologically relevant ones. After the first 24 hrs of transfection, the medium was replaced with DMEM with 10% FBS and after an additional 24 hrs, the cells were collected for analysis. RNA was extracted using the Direct-zol RNA kit and an in-column DNase digestion step was performed at room temperature for 15 minutes. The po

INSTRUMENT(S): Illumina Genome Analyzer II

ORGANISM(S): Homo sapiens

SUBMITTER: Gregor Rot 

PROVIDER: E-MTAB-4732 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress