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Abstract Although long-read single-cell RNA isoform sequencing (scISO-Seq) can reveal alternative RNA splicing in individual cells, it suffers from a low read throughput. Here, we introduce HIT-scISOseq, a method that removes most artifact cDNAs and concatenates multiple cDNAs for PacBio circular consensus sequencing (CCS) to achieve high-throughput and high-accuracy single-cell RNA isoform sequencing. HIT-scISOseq can yield >10 million high-accuracy long-reads in a single PacBio Sequel II SMRT Cell 8M. We also report the development of scISA-Tools that demultiplex HIT-scISOseq concatenated reads into single-cell cDNA reads with >99.99% accuracy and specificity. We apply HIT-scISOseq to characterize the transcriptomes of 3375 corneal limbus cells and reveal cell-type-specific isoform expression in them. HIT-scISOseq is a high-throughput, high-accuracy, technically accessible method and it can accelerate the burgeoning field of long-read single-cell transcriptomics. 

Configuration space complexity makes the big-data software systems hard to configure well. Consider Hadoop, with over nine hundred parameters, developers often just use the default configurations provided with Hadoop distributions. The opportunity costs in lost performance are significant. Popular learning-based approaches to auto-tune software does not scale well for big-data systems because of the high cost of collecting training data. We present a new method based on a combination of Evolutionary Markov Chain Monte Carlo (EMCMC)} sampling and cost reduction techniques tofind better-performing configurations for big data systems. For cost reduction, we developed and experimentally tested and validated two approaches: using scaled-up big data jobs as proxies for the objective function for larger jobs and using a dynamic job similarity measure to infer that results obtained for one kind of big data problem will work well for similar problems. Our experimental results suggest that our approach promises to improve the performance of big data systems significantly and that it outperforms competing approaches based on random sampling, basic genetic algorithms (GA), and predictive model learning. Our experimental results support the conclusion that our approach strongly demonstrates the potential toimprove the performance of big data systems significantly and frugally.