Homo sapiens. Homo sapiens

We demonstrate the use of 5’-Acrydite oligonucleotides to copolymerize single cell DNA or RNA into balls of acrylamide gel (BAG). Combining this step with pool and split techniques for creating barcodes yields a method with advantages in cost and scalability, efficient use of samples, depth of coverage, and ease of operation. We successfully perform DNA copy number profiling on mixtures of cell lines, nuclei from frozen prostate tumors, and biopsy washes. As applied to RNA the method has high capture efficiency of transcripts and sufficient consistency to distinguish clearly the expression patterns of two cell lines. Using varietal tags to achieve sequence error-correction, we show extremely low levels of cross-contamination by tracking source-specific SNVs. The method is readily modified, and we will discuss its future improvements and diverse applications.

我们展示了利用5’-Acrydite寡核苷酸将单细胞DNA或RNA共聚为丙烯酰胺凝胶微球（BAG）的方法。将该步骤与用于构建分子条形码的池化-拆分技术相结合，所得到的方法在成本、可扩展性、样本高效利用、测序覆盖深度以及操作便捷性上均具备显著优势。我们已成功在细胞系混合物、冷冻前列腺肿瘤组织的细胞核以及活检灌洗液样本中完成了DNA拷贝数谱分析。将该方法应用于RNA分析时，其对转录本的捕获效率极高，且一致性足以清晰区分两种细胞系的表达谱。通过引入样本特异性标签实现测序错误校正，我们通过追踪来源特异性单核苷酸变异（Single Nucleotide Variants，SNVs），证实该方法的交叉污染水平极低。该方法具备良好的可改造性，我们还将讨论其未来的改进方向与多样化应用场景。