Next Generation Sequencing Facilitates Quantitative Analysis of MM cell line Transcriptomes

We discovered that the activated-leukocyte-cell-adhesion-molecule (ALCAM) regulated myelomagenesis. ALCAM was a negative regulator of myeloma clonogenicity. To determine what signalling pathways are affected by ALCAM in MM cells, we established CTR-KD and ALCAM-KD RPMI8226 MM cell lines. Hoechst33342 staining was performed on CTR-KD and ALCAM-KD RPMI8226 MM cell. Then flow cytometry was used to separate MP and SP cells of the two types of cells. Total RNA of each sample was extracted using TRIzol Reagent (Invitrogen). Next generation sequencing library preparations were constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina (Vazyme, Cat No. NR603, China).RNA-seq was performed by GENEWIZ (Suzhou, China) on an Illumina Hiseq platform. Examination of mRNA profiles in CTR-KD MP, CTR-KD SP, ALCAM-KD MP and ALCAM-KD SP RPMI8226 cell.