Single cell libraries were prepared from PBMCs or toRNA from PBMCs using the QIAseq FX Single Cell RNA Library Kit and sequenced on NextSeq. Plotted is the % of reads that map to different RNA biotypes.
Single Cell RNA-Libraries from PBMCs and HeLa Cells were generated using QIAseq FX Single Cell RNA Library kit und a kit from Supplier C/I. Plotted are the percentage of reads that map to linc RNA detected in PBMC and HeLa preparations. QIAseq detects a significantly higher percentage of long regulatory RNAs compared to Supplier C/I.
4 Individual HeLa cells were isolated from the same cell culture and libraries were prepared with either the QIAseq FX Single Cell RNA Library Kit or a competing workflow. After sequencing to equal depth, quality control, alignment, and TPKM calculation, pairwise comparisons of the number of common transcripts detected with >1 TPKM divided by sum of all transcripts in both preps were made between all tested cells. The graphs represent mean of 6 pairwise comparisons with SD.
Single Cell RNA-Libraries from PBMCs were generated using QIAseq FX Single Cell RNA Library kit und a Kit from Supplier C/I. Libraries were sequenced on Illumina NextSeq. Plotted is the % of duplicates, that was obtained from the Fast QC report of the sequenced libraries.
The QIAseq FX Single Cell RNA Library Kit detects a greater number of transcripts at the same sequencing depth. To account for cell-to-cell differences in transcript abundance, libraries were produced from 100 pg of reference RNA from PBMCs. After sequencing, quality control and mapping, annotated transcripts with >1 TPM were quantified from either the full dataset or rarified sub-fractions. Saturation curves are from different sample preparation methods. Each point on the curve was generated by randomly selecting a number of raw reads from each sample library and then using the same alignment pipeline to call genes with mean TPM>1.