GSM8107922: HI3; Mus musculus; RNA-Seq
RNeasy Micro Kit Protocol Sequencing libraries were generated using Takara's SMARTR ZapR v2-based protocol; briefly, 4 ng of each RNA sample were used (corresponding to half of the reaction volume). Full volume was used for the 8 samples with the lowest concentration. RNAs were fragmented for 4 minutes, followed by first-strand cDNA synthesis using the SMARTR Pico v2 protocol. TruSeq-Singular unique dual index (UDI) sequencing adapters were added by PCR (5 cycles). Library fragments corresponding to rRNA molecules were depleted by cleaving with the ZapR v2 and mammalian-specific R-probe v2 kit at full volume, and the second round of PCR was performed using the Singular qPCR primers for 15 cycles. Library sizes were quantified and verified by QPCR and on a Fragment Analyzer before loading for sequencing on a Singular G4 instrument in a 50-base paired-end configuration.