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Method Article
Aviv Regev
Broad Institute
Corresponding Author
Feng Zhang
Broad Institute
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
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 as adult brain. In the adult brain the harsh enzymatic dissociation harms the integrity of the cells and their RNA, and biases toward easily dissociated cell types, and is restricted to young animals.
We developed DroNc-seq, a droplet microfluidic and DNA barcoding technique for analysis of RNA profiles 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.
Keywords
single nuclei, high throughput, RNA-seq, droplet, microfluidics, DNA barcode, frozen, RNAlater, tissue, hippocampus, PFC
N.H., A.B., I.A.D., D.A.W., F.Z. and A.R. are inventors on international patent application PCT/US16/59239 filed by Broad Institute, Harvard and MIT, relating to this protocol. The authors declare no competing financial interests.
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Associated Publications
Massively parallel single-nucleus RNA-seq with DroNc-seq
by Naomi Habib, Inbal Avraham-Davidi, Anindita Basu, +11
Nature Methods (29 August, 2017)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Aviv Regev
Broad Institute
Corresponding Author
Feng Zhang
Broad Institute
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 29 Aug, 2017
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
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 as adult brain. In the adult brain the harsh enzymatic dissociation harms the integrity of the cells and their RNA, and biases toward easily dissociated cell types, and is restricted to young animals.
We developed DroNc-seq, a droplet microfluidic and DNA barcoding technique for analysis of RNA profiles 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.
N.H., A.B., I.A.D., D.A.W., F.Z. and A.R. are inventors on international patent application PCT/US16/59239 filed by Broad Institute, Harvard and MIT, relating to this protocol. The authors declare no competing financial interests.
Loading...
Associated Publications
Massively parallel single-nucleus RNA-seq with DroNc-seq
by Naomi Habib, Inbal Avraham-Davidi, Anindita Basu, +11
Nature Methods (29 August, 2017)