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Method Article
DroNc-seq step-by-step
Aviv Regev
Broad Institute
Corresponding Author
Feng Zhang
Broad Institute
Corresponding Author
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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
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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)
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This is a preprint. It has not completed peer review.
Method Article
DroNc-seq step-by-step
Aviv Regev
Broad Institute
Corresponding Author
Feng Zhang
Broad Institute
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
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.
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)