PDF(Pubmed)
Abstract：
Here, we present a modification of single-cell tagged reverse transcription protocol to study gene expression on a single-cell level or with limited RNA input. We describe different enzymes for reverse transcription and cDNA amplification, modified lysis buffer, and additional clean-up steps before cDNA amplification. We also detail an optimized single-cell RNA sequencing method for handpicked single cells, or tens to hundreds of cells, as input material to study mammalian preimplantation development. For complete details on the use and execution of this protocol, please refer to Ezer et al.1.
摘要：
这里,我们提出了一个修改的单细胞标记的逆转录方案，以研究在单细胞水平或有限的RNA输入的基因表达。我们描述了逆转录和cDNA扩增的不同酶，改良的裂解缓冲液，和cDNA扩增前的额外清理步骤。我们还详细介绍了一种针对人工挑选的单细胞的优化的单细胞RNA测序方法，或数十到数百个细胞，作为研究哺乳动物植入前发育的输入材料。有关此协议的使用和执行的完整详细信息，PleaserefertoEzeretal.1.
