DroNc-seq step-by-step v1

Currently, most single cell protocols require the preparation of a single cell suspension from fresh tissue, a major roadblock to clinical deployment, to archived materials and to certain tissues such asadult brain. In the adultbrainthe harsh enzymatic dissociation harms the integrity of the cells and theirRNA, and biases toward easily dissociated cell types, and is restricted to young animals. We developed DroNc-seq, a droplet microfluidic andDNAbarcoding technique for analysis ofRNAprofiles of single nuclei from fresh, frozen or lightly fixed tissues at high throughput and low cost. The utility of DroNc-Seq lies in working with hard-to-dissociate, frozen and/or archived tissues. To demonstrate the utility of this technique, we sequenced over 39 thousand nuclei from mouse and human archived brain samples, including post-mortem human brain tissue from GTEx project.