title: Single Cell Total RNA Sequencing through Isothermal Amplification in Picoliter-Droplet Emulsion.
abstract: Prevalent single cell RNA amplification and sequencing chemistries mainly focus on polyadenylated RNAs in eukaryotic cells by using oligo(dT) primers for reverse transcription. We develop a new RNA amplification method, "easier-seq", to reverse transcribe and amplify the total RNAs, both with and without polyadenylate tails, from a single cell for transcriptome sequencing with high efficiency, reproducibility, and accuracy. By distributing the reverse transcribed cDNA molecules into 1.5 × 105aqueous droplets in oil, the cDNAs are isothermally amplified using random primers in each of these 65-pL reactors separately. This new method greatly improves the ease of single-cell RNA sequencing by reducing the experimental steps. Meanwhile, with less chance to induce errors, this method can easily maintain the quality of single-cell sequencing. In addition, this polyadenylate-tail-independent method can be seamlessly applied to prokaryotic cell RNA sequencing.
authors: Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, and Beijing Advanced Innovation Center for Genomics (ICG), Peking University , Beijing 100871, China.
authors: Yusi
authors: Y
authors: Fu
authors: Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, and Beijing Advanced Innovation Center for Genomics (ICG), Peking University , Beijing 100871, China.
authors: He
authors: H
authors: Chen
authors: Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, and Beijing Advanced Innovation Center for Genomics (ICG), Peking University , Beijing 100871, China.|College of Engineering, Peking University , Beijing 100871, China.
authors: Lu
authors: L
authors: Liu
authors: Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, and Beijing Advanced Innovation Center for Genomics (ICG), Peking University , Beijing 100871, China.|College of Engineering, Peking University , Beijing 100871, China.|Peking-Tsinghua Center for Life Sciences, Peking University , Beijing 100871, China.
authors: Yanyi
authors: Y
authors: Huang
mesh_terms: D004655:Emulsions / Q000737:chemistry; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010316:Particle Size; D012313:RNA / Q000032:analysis* / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods*; D059010:Single-Cell Analysis / Q000379:methods*; D013499:Surface Properties
chemical_list: D004655:Emulsions; D012313:RNA