| Chapter title |
Multiplexed Targeted Sequencing for Oxford Nanopore MinION: A Detailed Library Preparation Procedure
|
|---|---|
| Chapter number | 4 |
| Book title |
Next Generation Sequencing
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7514-3_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7512-9, 978-1-4939-7514-3
|
| Authors |
Timokratis Karamitros, Gkikas Magiorkinis |
| Abstract |
MinION is a small form factor sequencer recently retailed by Oxford Nanopore technologies. This lighter-sized USB3.0-interfaced device uses innovative nanotechnology to generate extra-long reads from libraries prepared using only standard molecular biology lab equipment. The flexibility and the portability of the platform makes it ideal for point-of-interest and real-time surveillance applications. However, MinION's limited capacity is not enough for the study of specific targets within larger genomes. Apart from just PCR-amplifying regions of interest, the capture of long reads spanning the edges of known-unknown genomic regions is of great importance for structural studies, such as the identification of mobile elements' integrations sites, bridging over low complexity repetitive regions etc.In this study, using MinION-kit-included and commercially available reagents, we have developed an easy and versatile wet-lab procedure for the targeted enrichment of MinION libraries, capturing DNA fragments of interest before the ligation of the sensitive MinION sequencing-adapters. This method allows for simultaneous target-enrichment and barcode-multiplexing of up to 12 libraries, which can be loaded in the same sequencing run. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 5 | 12% |
| United States | 5 | 12% |
| Australia | 4 | 10% |
| Netherlands | 3 | 7% |
| Germany | 2 | 5% |
| South Africa | 2 | 5% |
| Sweden | 2 | 5% |
| Spain | 2 | 5% |
| Taiwan | 1 | 2% |
| Other | 2 | 5% |
| Unknown | 14 | 33% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 28 | 67% |
| Members of the public | 14 | 33% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 109 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 21 | 19% |
| Researcher | 21 | 19% |
| Student > Master | 11 | 10% |
| Student > Bachelor | 9 | 8% |
| Professor | 5 | 5% |
| Other | 14 | 13% |
| Unknown | 28 | 26% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 34 | 31% |
| Agricultural and Biological Sciences | 17 | 16% |
| Neuroscience | 5 | 5% |
| Immunology and Microbiology | 4 | 4% |
| Engineering | 3 | 3% |
| Other | 11 | 10% |
| Unknown | 35 | 32% |