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.
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.
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.
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.
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.