Anthropology & Human Genomics, Faculty of Biology, Ludwig-Maximilians University, Großhaderner Str. 2, 82152, Martinsried, Germany.
Graduate School of Systemic Neurosciences, Faculty of Biology, Ludwig-Maximilians University, Martinsried, Germany.
Genome Biol. 2022 Mar 31;23(1):88. doi: 10.1186/s13059-022-02660-8.
Cost-efficient library generation by early barcoding has been central in propelling single-cell RNA sequencing. Here, we optimize and validate prime-seq, an early barcoding bulk RNA-seq method. We show that it performs equivalently to TruSeq, a standard bulk RNA-seq method, but is fourfold more cost-efficient due to almost 50-fold cheaper library costs. We also validate a direct RNA isolation step, show that intronic reads are derived from RNA, and compare cost-efficiencies of available protocols. We conclude that prime-seq is currently one of the best options to set up an early barcoding bulk RNA-seq protocol from which many labs would profit.
通过早期条形码进行具有成本效益的文库生成一直是推动单细胞 RNA 测序的核心。在这里，我们优化并验证了 prime-seq，这是一种早期条形码批量 RNA-seq 方法。我们发现它与标准的批量 RNA-seq 方法 TruSeq 的性能相当，但由于文库成本便宜近 50 倍，因此效率提高了四倍。我们还验证了直接 RNA 分离步骤，表明内含子读数来自 RNA，并比较了现有方案的成本效率。我们得出结论，prime-seq 是目前建立早期条形码批量 RNA-seq 协议的最佳选择之一，许多实验室将从中受益。
