mirror of
https://github.com/scinote-eln/scinote-web.git
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1014 lines
No EOL
40 KiB
JSON
1014 lines
No EOL
40 KiB
JSON
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"text": "Final libraries are put through quality control and high-throughput sequencing.",
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"text": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body>\n<p> The RAD library was prepared as originally described in <a href=\"https://www.ncbi.nlm.nih.gov/pubmed/18852878/\" target=\"_blank\" rel=\"noopener\">article</a>.<br><br>At least <strong>30</strong> specimens need to be chosen.<br><br><strong>Each sample is digested at 37°C for 40 min with restriction enzyme:</strong></p>\n<ul>\n<li>\n<strong>6 U</strong> <em>restriction enzyme</em> per μg genomic DNA in <strong>1×</strong> Reaction Buffer 4 at a final concentration of about <strong>1 μg</strong> DNA per <strong>50 μL</strong> reaction volume</li>\n<li>The samples are heat-inactivated at <strong>65°C</strong> for <strong>20 min</strong>.</li>\n</ul>\n<br><strong>Individual-specific P1 adapters, each with a unique 5 or 7 bp barcode is ligated to the digested DNA at 22°C for 15 min by adding:</strong><br><br>\n<ul>\n<li>0.6 μL (DNA samples) 100 nmol/L P1 adapter</li>\n<li>0.15 μL 100 mmol/L rATP </li>\n<li>0.25 μL 10× Reaction Buffer 2</li>\n<li>0.125 μL T4 ligase </li>\n<li>reaction volumes made up to 15 μL with nuclease-free water </li>\n</ul>\n<br>After heat-inactivation, the ligation reactions are slowly cooled to room temperature then combined in appropriate multiplex pools.</body></html>"
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"text": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body>\n<p>Reads of low quality must be discarded. <br>Retained reads are sorted into loci and genotypes using software developed for that purpose. It will assigns loci using a likelihood-based algorithm to separate actual SNPs from SNPs likely to have arisen from sequencing error. <br><br><strong>Parameters:</strong><br>\n</p>\n<ul>\n<li>De novo assembly parameters.</li>\n<li>Minimum stack depth of 5.</li>\n<li>Maximum of 2 mismatches.</li>\n<li>No more than 1 mismatch between alleles.</li>\n</ul>\n</body></html>"
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"text": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body><p>Sequencing data from filtered Informative RAD markers is combined into a single alignment of alleles in RAD library construction. Data analysis is carried out using R and an associated R/<em>adegenet</em> package for Principal Component Analysis (PCA) and Discriminant Analysis of Principal Components (DAPC).<em><strong> PCA creates simplified models of the total variation within the dataset.</strong> <strong>DAPC identifies clusters of genetically related individuals.</strong></em></p></body></html>"
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|
||
"text": "Add a ½ volume of 5 M NaCl to the sample and mix gently by inversion.",
|
||
"checked": false,
|
||
"checklist_id": 959,
|
||
"created_at": "2018-12-21T09:54:21.496Z",
|
||
"updated_at": "2018-12-21T09:54:21.496Z",
|
||
"created_by_id": null,
|
||
"last_modified_by_id": null,
|
||
"position": 0
|
||
},
|
||
{
|
||
"id": 3944,
|
||
"text": "Then, add 3 volumes of cold 95% ethanol and mix gently by inversion.",
|
||
"checked": false,
|
||
"checklist_id": 959,
|
||
"created_at": "2018-12-21T09:54:21.503Z",
|
||
"updated_at": "2018-12-21T09:54:21.503Z",
|
||
"created_by_id": null,
|
||
"last_modified_by_id": null,
|
||
"position": 1
|
||
},
|
||
{
|
||
"id": 3945,
|
||
"text": "Place the tubes into a -20°C freezer and incubate for 1 h.",
|
||
"checked": false,
|
||
"checklist_id": 959,
|
||
"created_at": "2018-12-21T09:54:21.509Z",
|
||
"updated_at": "2018-12-21T09:54:21.509Z",
|
||
"created_by_id": null,
|
||
"last_modified_by_id": null,
|
||
"position": 2
|
||
},
|
||
{
|
||
"id": 3946,
|
||
"text": "After incubation, centrifuge the Falcon tube for 10 min at 5000 × g to pellet the DNA.",
|
||
"checked": false,
|
||
"checklist_id": 959,
|
||
"created_at": "2018-12-21T09:54:21.516Z",
|
||
"updated_at": "2018-12-21T09:54:21.516Z",
|
||
"created_by_id": null,
|
||
"last_modified_by_id": null,
|
||
"position": 3
|
||
},
|
||
{
|
||
"id": 3947,
|
||
"text": "Carefully decant away the supernatant and wash the DNA pellet with 3 mL of 70% ethanol.",
|
||
"checked": false,
|
||
"checklist_id": 959,
|
||
"created_at": "2018-12-21T09:54:21.522Z",
|
||
"updated_at": "2018-12-21T09:54:21.522Z",
|
||
"created_by_id": null,
|
||
"last_modified_by_id": null,
|
||
"position": 4
|
||
},
|
||
{
|
||
"id": 3948,
|
||
"text": "Gently swirl the solution and centrifuge again for 10 min at 5000 × g.",
|
||
"checked": false,
|
||
"checklist_id": 959,
|
||
"created_at": "2018-12-21T09:54:21.529Z",
|
||
"updated_at": "2018-12-21T09:54:21.529Z",
|
||
"created_by_id": null,
|
||
"last_modified_by_id": null,
|
||
"position": 5
|
||
},
|
||
{
|
||
"id": 3949,
|
||
"text": "Carefully decant the supernatant and air-dry DNA pellet for 15 min at room temperature. Once dried, suspend DNA in 200 μL of TE buffer.",
|
||
"checked": false,
|
||
"checklist_id": 959,
|
||
"created_at": "2018-12-21T09:54:21.535Z",
|
||
"updated_at": "2018-12-21T09:54:21.535Z",
|
||
"created_by_id": null,
|
||
"last_modified_by_id": null,
|
||
"position": 6
|
||
}
|
||
]
|
||
},
|
||
"position": 1
|
||
}
|
||
]
|
||
}
|
||
]
|
||
}
|
||
],
|
||
"results": [
|
||
|
||
]
|
||
}
|
||
],
|
||
"my_module_groups": [
|
||
{
|
||
"id": 1167,
|
||
"created_at": "2018-12-17T07:51:25.142Z",
|
||
"updated_at": "2018-12-17T07:51:25.142Z",
|
||
"created_by_id": 202,
|
||
"experiment_id": 433
|
||
}
|
||
]
|
||
} |