As new NGS platforms become available, studies are required to shift the sequencings plans. This is a comparison of the same series of libraries run on older HiSeq systems (HS2500 and HS4000) and the newer Nextseq and Novaseq platforms. These were TruSeq stranded mRNA. Pools of 4 libraries were run on the HiSeq/ paired end/ 50x. The same pools were then pooled into larger pools (pools of pools) to be run on the larger platforms.