GSM4494797: APP-6m-sR_3; Mus musculus; RNA-Seq
Trizol RNA extraction: 1mL trizol added to pelleted cells, dissolved 5min. 200uL chloroform added to trizol treated cells, incubate 2min. Centrifuged at 12 000xg 4C. Aqueous layer treated with 500uL isopropanol, incubate -20C 1hour, centrifuged at 12 000xg 10min 4C, pellet washed with 75% ethanol and centrifuged at 7 500xg 5min 4C. Pellet dried 1min, resuspended in 30uL pure H2O Libraries prepared as described before (Zovoilis et al, Cell 2016) and in the manuscript that accompanies the current submission. Enrichemnt of short RNAs was done using a  modified protocol for the MiRvana size selection kit. Short-RNA-seq library performed using NEBNext Multiplex Small Library Prep E7560S and a customized bead cleaning approach to get rid of very small fragments (miRNAs) and adapter dimers. Long-RNA-seq library performed after fragmentation and ribosomal RNA depletion with NEBNext rRNA depletion kit using  NEBNext Ultra RNA Library Prep Kit for Illumina (E7530)
GEO
GSM4494797
GEO Accession
GSM4494797