| Chapter title |
The Use of High-Throughput Sequencing for the Study and Diagnosis of Plant Viruses and Viroids in Pollen
|
|---|---|
| Chapter number | 10 |
| Book title |
Viral Metagenomics
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7683-6_10 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7682-9, 978-1-4939-7683-6
|
| Authors |
Kris De Jonghe, Annelies Haegeman, Yoika Foucart, Martine Maes |
| Abstract |
This protocol details the wet lab preparation, extraction of fruit pollen samples, and analysis of the sequencing data following Illumina NextSeq small and total RNA sequencing. The protocol was developed for virus and viroid detection using NGS sequencing and was based on the results of a comparison between different extraction methods followed by yield, RNA purity, and integrity assessment. Moreover, the advantage of an additional ribosomal (r)RNA depletion step to the total RNA extraction protocol was evaluated. The smallRNA procedure is the preferred method of choice. If the total RNA protocol is chosen, the use of the mirVana kit followed by an rRNA depletion step is the best option. The library preparation and sequencing steps were outsourced. As a final step in the data analysis, the VirusDetect software was used to detect the viruses and viroids in the pollen samples. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 17 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 7 | 41% |
| Student > Ph. D. Student | 2 | 12% |
| Student > Bachelor | 2 | 12% |
| Lecturer | 1 | 6% |
| Student > Doctoral Student | 1 | 6% |
| Other | 1 | 6% |
| Unknown | 3 | 18% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 9 | 53% |
| Biochemistry, Genetics and Molecular Biology | 5 | 29% |
| Unknown | 3 | 18% |