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Single-Cell mRNA-Seq Using the Fluidigm C1 System and Integrated Fluidics Circuits.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1783, pp. 193-207. - Publication Year :
- 2018
-
Abstract
- Single-cell mRNA-seq is a valuable tool to dissect expression profiles and to understand the regulatory network of genes. Microfluidics is well suited for single-cell analysis owing both to the small volume of the reaction chambers and easiness of automation. Here we describe the workflow of single-cell mRNA-seq using C1 IFC, which can isolate and process up to 96 cells. Both on-chip procedure (lysis, reverse transcription, and preamplification PCR) and off-chip sequencing library preparation protocols are described. The workflow generates full-length mRNA information, which is more valuable compared to 3' end counting method for many applications.
- Subjects :
- DNA, Complementary biosynthesis
DNA, Complementary genetics
Gene Library
Humans
Microfluidic Analytical Techniques methods
RNA, Messenger genetics
Real-Time Polymerase Chain Reaction
Gene Expression Profiling methods
High-Throughput Nucleotide Sequencing methods
Microfluidic Analytical Techniques instrumentation
RNA, Messenger analysis
Sequence Analysis, RNA methods
Single-Cell Analysis methods
Transcriptome
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1783
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 29767364
- Full Text :
- https://doi.org/10.1007/978-1-4939-7834-2_10