GSM2052066: 22dAscl1-iN_iN7_16; Mus musculus; RNA-Seq
Homozygous TauEGFP knock-in mice21 were purchased from the Jackson Laboratories and bred with C57BL/6 mice (Taconic) to generate TauEGFP heterozygous embryos. MEFs were isolated from E14.5 embryos using a dissection microscope (Leica). Tail tips were sliced into small pieces, trypsinized and plated to derive fibroblast cultures. All fibroblasts were expanded for three passages before being used for experiments. Complementary DNAs for candidate genes were cloned into doxycycline-inducible lentiviral vectors. Clonal Ascl1-inducible MEFs were derived as described by Wapinski et al., Cell 2013. Cells were loaded into the Fluidigm C1 microfluidic platform, where single cells were captured. Lysis of single cells, reverse-transcription of mRNA into cDNA as well as preamplification of cDNA occured within the microfluidic device using reagents provided by Fluidigm as well as the SMARTer Ultra Low RNA kit for Illumina (Clontech). External RNA spike-in transcripts (ERCC spike-in Mix, Ambion) were added to all single cell lysis reactions at a dilution of 1:40,000 (and 1:4,000 in case of 5d experiment). Libraries were prepared using Illumina Nextera XT kit per illumina's protocols.
GEO Accession
GSM2052066