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Single-Cell Semiconductor Sequencing
- Source :
- Methods in Molecular Biology ISBN: 9781627035552
- Publication Year :
- 2013
- Publisher :
- Humana Press, 2013.
-
Abstract
- RNA-seq or transcriptome analysis of individual cells and small-cell populations is essential for virtually any biomedical field. It is especially critical for developmental, aging, and cancer biology as well as neuroscience where the enormous heterogeneity of cells present a significant methodological and conceptual challenge. Here we present two methods that allow for fast and cost-efficient transcriptome sequencing from ultra-small amounts of tissue or even from individual cells using semiconductor sequencing technology (Ion Torrent, Life Technologies). The first method is a reduced representation sequencing which maximizes capture of RNAs and preserves transcripts' directionality. The second, a template-switch protocol, is designed for small mammalian neurons. Both protocols, from cell/tissue isolation to final sequence data, take up to 4 days. The efficiency of these protocols has been validated with single hippocampal neurons and various invertebrate tissues including individually identified neurons within a simpler memory-forming circuit of Aplysia californica and early (1-, 2-, 4-, 8-cells) embryonic and developmental stages from basal metazoans.
- Subjects :
- Neurons
Aging
Genome
Base Sequence
biology
Sequence Analysis, RNA
Gene Expression Profiling
Cell
Computational biology
Ion semiconductor sequencing
biology.organism_classification
Hippocampus
Embryonic stem cell
Molecular biology
Article
Transcriptome
Gene expression profiling
medicine.anatomical_structure
Aplysia
medicine
Animals
Directionality
Subjects
Details
- ISBN :
- 978-1-62703-555-2
- ISBNs :
- 9781627035552
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology ISBN: 9781627035552
- Accession number :
- edsair.doi.dedup.....51c31ed2665e9788a1e7617278814e7b