GSM4680640: ABC-imKO3 Naïve rep1 line 1; Mus musculus; RNA-Seq
Microglia cells were sorted directtly into lysis buffer buffer containing Triton X-100, RNase Inhibitor, and Oligo(dT)30-VN and vortexed. Then lysates were hybridized with oligo(dT)+ to the poly(A) tails of the mRNA. Reagents for reverse transcription were added to construct cDNA libraries following addition of reagents for PCR amplification (qPCR was not performed at this point). Libraries were quantified and QC was performed using TapeStation high sensitivity D5000 screentape in addition to Qubit double stranded high sensitivity assay. All samples were adjusted to 1ng of cDNA for input into NexteraXT protocol. QC check was performed with TapeStation high sensitivity D1000 screen tape in addition to Qubit double stranded high sensitivity assay. The samples were subjected to qPCR and pooling and loaded onto the NovaSeq for paired-end 50x50 reads using NovaSeq S1 100 cycle kit. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM4680640