Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq. Bagnoli, J. W., Ziegenhain, C., Janjic, A., Wange, L. E., Vieth, B., Parekh, S., Geuder, J., Hellmann, I., & Enard, W. Nature Communications, 9:2937, July, 2018.
Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq [link]Paper  doi  abstract   bibtex   
Single-cell RNA sequencing (scRNA-seq) has emerged as a central genome-wide method to characterize cellular identities and processes. Consequently, improving its sensitivity, flexibility, and cost-efficiency can advance many research questions. Among the flexible plate-based methods, single-cell RNA barcoding and sequencing (SCRB-seq) is highly sensitive and efficient. Here, we systematically evaluate experimental conditions of this protocol and find that adding polyethylene glycol considerably increases sensitivity by enhancing cDNA synthesis. Furthermore, using Terra polymerase increases efficiency due to a more even cDNA amplification that requires less sequencing of libraries. We combined these and other improvements to develop a scRNA-seq library protocol we call molecular crowding SCRB-seq (mcSCRB-seq), which we show to be one of the most sensitive, efficient, and flexible scRNA-seq methods to date., Single-cell RNA-barcoding and sequencing is an efficient, genome-wide method to characterize cellular identities. Here the authors systematically evaluate the protocol and develop molecular crowding SCRB-seq with improved sensitivity and cost-efficiency.
@article{bagnoli_sensitive_2018,
	title = {Sensitive and powerful single-cell {RNA} sequencing using {mcSCRB}-seq},
	volume = {9},
	issn = {2041-1723},
	url = {https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6062574/},
	doi = {10.1038/s41467-018-05347-6},
	abstract = {Single-cell RNA sequencing (scRNA-seq) has emerged as a central genome-wide method to characterize cellular identities and processes. Consequently, improving its sensitivity, flexibility, and cost-efficiency can advance many research questions. Among the flexible plate-based methods, single-cell RNA barcoding and sequencing (SCRB-seq) is highly sensitive and efficient. Here, we systematically evaluate experimental conditions of this protocol and find that adding polyethylene glycol considerably increases sensitivity by enhancing cDNA synthesis. Furthermore, using Terra polymerase increases efficiency due to a more even cDNA amplification that requires less sequencing of libraries. We combined these and other improvements to develop a scRNA-seq library protocol we call molecular crowding SCRB-seq (mcSCRB-seq), which we show to be one of the most sensitive, efficient, and flexible scRNA-seq methods to date., Single-cell RNA-barcoding and sequencing is an efficient, genome-wide method to characterize cellular identities. Here the authors systematically evaluate the protocol and develop molecular crowding SCRB-seq with improved sensitivity and cost-efficiency.},
	urldate = {2022-02-04},
	journal = {Nature Communications},
	author = {Bagnoli, Johannes W. and Ziegenhain, Christoph and Janjic, Aleksandar and Wange, Lucas E. and Vieth, Beate and Parekh, Swati and Geuder, Johanna and Hellmann, Ines and Enard, Wolfgang},
	month = jul,
	year = {2018},
	pmid = {30050112},
	pmcid = {PMC6062574},
	pages = {2937},
}

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