Prevalent single cell RNA amplification and sequencing chemistries mainly focus on polyadenylated RNAs in eukaryotic cells by using oligo(dT) primers for reverse transcription. Researchers from Peking University have developed 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 1.5×105 aqueous droplets in oil, the cDNAs are isothermally amplified using random primers in each of these 65-picoliter reactors separately. This new method greatly improves the ease of single-cell RNA sequencing by reducing the experimental steps. 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.
Single cell total RNA sequencing through isothermal amplification in picoliter-droplet emulsion
Fu Y, Chen H, Liu L, Huang Y. (2016) Single cell total RNA sequencing through isothermal amplification in picoliter-droplet emulsion. Anal Chem [Epub ahead of print]. [abstract]