Chapter title |
Preparation of Splicing Competent Nuclear Extract from Mammalian Cells and In Vitro Pre-mRNA Splicing Assay
|
---|---|
Chapter number | 2 |
Book title |
mRNA Processing
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-7204-3_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7203-6, 978-1-4939-7204-3
|
Authors |
Maliheh Movassat, Hossein Shenasa, Klemens J. Hertel |
Abstract |
The ability to perform in vitro splicing assays has paved the way for in-depth studies of the mechanisms and machinery involved in the process of splicing. The in vitro splicing assay is a valuable experimental approach that combines the complexity of the spliceosome and regulatory systems with the flexibility of performing endless splicing and alternative splicing reactions. Through the use of crude nuclear extract and radiolabeled pre-mRNA, spliced mRNAs can be visualized using autoradiography for downstream analysis. This chapter describes the necessary steps to perform an in vitro splicing reaction, including the generation of the key components necessary for the splicing reaction; nuclear extract. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 20 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 4 | 20% |
Student > Doctoral Student | 3 | 15% |
Professor | 2 | 10% |
Student > Ph. D. Student | 2 | 10% |
Student > Master | 2 | 10% |
Other | 3 | 15% |
Unknown | 4 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 8 | 40% |
Agricultural and Biological Sciences | 4 | 20% |
Chemical Engineering | 1 | 5% |
Immunology and Microbiology | 1 | 5% |
Materials Science | 1 | 5% |
Other | 0 | 0% |
Unknown | 5 | 25% |