Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq
| العنوان: | Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq |
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| المؤلفون: | Lucas E. Wange, Johannes W. Bagnoli, Wolfgang Enard, Johanna Geuder, Aleksandar Janjic, Swati Parekh, Beate Vieth, Ines Hellmann, Christoph Ziegenhain |
| المصدر: | Nature Communications, Vol 9, Iss 1, Pp 1-8 (2018) Nature Communications |
| بيانات النشر: | Nature Portfolio, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Sequence analysis, Computer science, Science, genetic processes, Cell, General Physics and Astronomy, Computational biology, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Library Protocol, Single-cell analysis, Complementary DNA, medicine, natural sciences, lcsh:Science, Polymerase, Multidisciplinary, Base Sequence, biology, Sequence Analysis, RNA, High-Throughput Nucleotide Sequencing, RNA, General Chemistry, 030104 developmental biology, medicine.anatomical_structure, biology.protein, lcsh:Q, Research questions, Single-Cell Analysis, Software |
| الوصف: | 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. |
| اللغة: | English |
| تدمد: | 2041-1723 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::713e8f80a7592fca6db998d81d92a0c4Test https://doaj.org/article/4a131646ab114fc18a5468c079161e41Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....713e8f80a7592fca6db998d81d92a0c4 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20411723 |
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