Figure 1: SEQC study design.
From: Cross-platform ultradeep transcriptomic profiling of human reference RNA samples by RNA-Seq
This figure was modified from b presented in the related research manuscript13. Similar to the MAQC-I benchmarks, well characterized RNA samples A and B were augmented by samples C and D comprised of A and B in known mixing ratios 3:1 and 1:3, respectively. These allow tests for titration consistency and the correct recovery of the known mixing ratios. Synthetic RNAs from the External RNA Control Consortium (ERCC) were both pre-added to samples A and B before mixing and also sequenced separately to assess dynamic range (samples E and F). Samples were distributed to independent sites for RNA-Seq library construction and profiling by Illumina’s HiSeq 2000 (3+4x) and Life Technologies’ SOLiD 5500 (3+1x). In addition to the replicate libraries A1…D4 at each site, for each platform, one vendor-prepared library A5…D5 was being sequenced at all three official sites, giving a total of 24 libraries. At each site, each library has a unique barcode sequence and all libraries were pooled before sequencing, so each lane was sequencing the same material, allowing a study of lane specific effects. Samples A and B were also sequenced by Roche 454 GS FLX at different sites with two runs each but no library replicates.
