GSM5031911: Wnt1Cre Mutant #4; Mus musculus; RNA-Seq
ENCDC were isolated from individual E11.5 stomachs and E13.5 colons by incubating fetal organs with collagenase and dispase in Ham's F12 media with 1% bovine serum albumin (37oC, 5% CO2 incubator, 20 minutes) and then triturating by pipetting up and down 15 times using a P1000 pipette tip before passing through a 30 μm strainer. Cells with TdTomato or EYFP fluorescence were purified by flow sorting (MoFlo Astrios high speed cell sorter, 100 μm nozzle), pelleted by centrifugation (500 x g), washed once with PBS, and frozen immediately at -80oC until analysis. RNA isolation, library preparation, and RNA-SEQ analysis were performed by University of Pennsylvania Next-Generation Sequencing Core. The libraries were made using the combination of the Clontech (now Takara) SMART (cDNA synthesis and amplification) and Illumina Nextera XT (cDNA to library) kits. Libraries were then sequenced on an Illumina HiSeq 4000, using Sequencer Softwares RTA: 2.7.7 and HCS: HiSeqCS:HD 3.4.0.38. To obtain enough reads, each sample was sequenced twice, and the results were combined together.
GEO
GSM5031911