GSM8284854: PyTCas9BROAD_FVB1 Rejected Cas9-BROAD Py2T Mammary fat pad tumor; Mus musculus; RNA-Seq
Individual tumor fragments were and homogenized using M-tubes (Miltenyi) and GentleMACS devices (Miltenyi) using the program RNA_001 and total RNA was isolated using the RNeasy kit (Qiagen). The TruSeq Stranded mRNA (Illumina) was used for library preparation as described. Briefly, total RNA samples (500 ng) were poly-A enriched and then reverse-transcribed into double-stranded cDNA. The cDNA samples were fragmented, end-repaired and adenylated before ligation of TruSeq adapters containing unique dual indices (UDI) for multiplexing. Fragments containing TruSeq adapters on both ends were selectively enriched by PCR. The quality and quantity of the enriched libraries were validated using the Fragment Analyzer (Agilent).