GSM9210848: Muscle.60mins.Control-5; Mus musculus; RNA-Seq
Followed the Rneasy Mini Kit for RNA extraction Ten microliters of undiluted total RNA was incubated with 10ul of NEB Dynabeads for polyaA+ RNA selection, eluted, and fragmented in 20ul. Illumina libraries were constructed using NEB Ultra II protocol; 5ul of the fragmented solution using half the amount of reagent and 1,2 PrD instead of enhancer in the ligation step, with 14 PCR cycles of final amplification and TS Singular index barcoding.Library sizes were quantified and verified by QPCR and on a Fragmen t Analyzer before loading for sequencing on a Singular G4 instrument in a 50-base paired-end configuration. Low-input samples were prepared by ribodepletion using the ZapR v2 protocol and mammalian-specific R-probe V2 kit at full volume (7.5 min fragmentation time).