GSM5552484: WMI31_C; Mus musculus; RNA-Seq
2,000 plasma cells were FACS-sorted into 75µL RLT buffer (QIAGEN, supplemented with BME) and frozen at -80°C. RNA was harvested using the RNeasy Micro kit (QIAGEN). Then cDNA was synthesized and amplified following the low-input RNA-Seq protocol from Rosales, Liang, Engel et al. (2019). cDNA was synthesized using SuperScriptII reverse transcriptase (Invitrogen), an Oligo-dT30VN and a TSO (template switching oligo); then cDNA was amplified using the KAPA HiFi polymerase (Roche) and the Oligo ISPCR as described. cDNA libraries were constructed using the Nextera XT DNA library preparation kit (Illumunia) with Tagmentation (Tn5 DNA insertion method) to incorporate sequencing indices (Nextera XT index kit, Illumina). Finally AMPure beads (Beckman Coulter) were employed to purify the library.