| Chapter title |
Manipulation of Oat Protoplasts for Transient Expression Assays
|
|---|---|
| Chapter number | 5 |
| Book title |
Oat
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6682-0_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6680-6, 978-1-4939-6682-0
|
| Authors |
Robyn Roberts, Jincan Zhang, Nicole Mihelich, Danielle Savino, Aurélie M. Rakotondrafara |
| Editors |
Sebastian Gasparis |
| Abstract |
Oat protoplasts are a useful and convenient system to study transient expression using whole cells. Nucleic acid can rapidly be introduced into live cells, and, depending on the assay, results can be collected the same day. Compared to plant tissue, oat cell suspension cultures provide a simple, high yielding, and consistent means to isolate protoplasts. Here, we describe how to generate an oat cell suspension culture from callus grown on solidified medium, and how to maintain the oat cells in cell suspension culture for protoplast preparation. Following the culturing procedure, we describe how to isolate oat protoplasts from cell suspension culture by enzymatic digestion of the cell walls and to transiently express nucleic acid (DNA or RNA) into the cells by electroporation. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 8 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 50% |
| Student > Master | 1 | 13% |
| Unknown | 3 | 38% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 4 | 50% |
| Agricultural and Biological Sciences | 1 | 13% |
| Social Sciences | 1 | 13% |
| Unknown | 2 | 25% |