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# Adding GO term and InterPro domain definition | ||
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Descriptions for GO terms and InterPro domains should be added before | ||
adding functional annotation. This step should be completed first. In | ||
the top menu click on 'Add' and select 'Functional Data'. | ||
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 | ||
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The GO descriptions can be obtained from the Gene Ontology Consortium's | ||
official website in OBO format [here](http://geneontology.org/page/download-ontology). | ||
InterPro domains and descriptions (called the **Entry list**) are found on EBI InterPro's download pages [here](https://www.ebi.ac.uk/interpro/download.html), | ||
decompress the .gz file prior to uploading. | ||
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Click the buttons on the page and select the corresponding files, next | ||
click 'Add functional data' to upload the files to your server and | ||
import them in the database. This process can take some time, do not | ||
close the browser window. | ||
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**Note: The existing tables will be cleared before adding the new | ||
definitions. Do not update this information if GO/InterPro data is | ||
already added to species !** |
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# Adding a new species and functional data | ||
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Adding a species requires multiple steps, follow the steps below for | ||
each species. | ||
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## Adding the species and sequences | ||
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On the 'Admin panel', under 'Add' select 'Species'. Fill in the | ||
full scientific name (or the name you wish to appear on the website) for | ||
the species and select a 'three letter code', that is unque for the | ||
species (we recommend a combination of genus and species cfr. **H**omo | ||
**sa**piens = hsa, note that while three characters is recommended it is | ||
not required, longer codes are possible). | ||
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Some visualizations require a color specific for each species, these can | ||
be entered using the controls below (clicking the box opens a color | ||
picker, there is no need to manually add in hex values). | ||
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Finally, select a fasta file with **coding sequences**, in the fasta | ||
headers the gene name (and only the name) needs to be present. These are | ||
the names genes will receive on the website. | ||
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``` | ||
>Gene1 | ||
ATG... | ||
>Gene2 | ||
ATG... | ||
``` | ||
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To upload the data and add the species to the database click 'Add species' | ||
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## Adding descriptions to sequences | ||
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In the 'Admin panel', under 'Add' -> 'Sequence descriptions'. | ||
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Select the **species** and a tab-delimited file containing on a single line the gene ID and | ||
description (example below). Click **Add descriptions** to upload the file and add the | ||
description to the database. | ||
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``` | ||
gene01 gene01 description | ||
gene02 gene02 description | ||
gene03 gene03 description | ||
... | ||
``` | ||
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Note: This step can be very time consuming if Whooshee indexing is enabled in your config! When | ||
setting up a database with multiple species, consider disabling indexing while building the DB and | ||
later enabling it + rebuilding the index (found under controls in the admin panel). | ||
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## Adding functional Annotation to sequences | ||
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**GO** terms can be imported from tab delimited files, containing 3 | ||
columns: gene name, GO label and the evidence tag. | ||
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``` | ||
Gene1 GO:0004621 IEA | ||
Gene1 GO:0004622 IEA | ||
Gene2 GO:0000227 ISS | ||
... | ||
``` | ||
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On the 'Admin panel', under 'Add' -> 'GO Genes' such a file can be | ||
uploaded, for a species. Additionally a description needs to be provided | ||
from where the GO terms in the file originate from (the source). | ||
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 | ||
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**InterPro** domains can be imported directly from [InterProScan](http://www.ebi.ac.uk/interpro/download.html) output. | ||
To do so, in the 'Admin panel', under 'Add' -> 'InterPro genes' select a species, select the file and click 'Add InterPro' | ||
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 |
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# Adding expression data | ||
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Expression data should be processed using [LSTrAP](https://github.molgen.mpg.de/proost/LSTrAP), | ||
this will generate the expression matrix, coexpression networks and | ||
clusters that can be directly imported into CoNekt. Note that in | ||
some cases additional files, containing meta information, need to be | ||
provided. | ||
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## Adding expression profiles | ||
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In the 'Admin panel', under 'Add' -> 'Expression profiles'. Select the | ||
species and the source (currently only LSTrAP expression matrices are supported). | ||
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Next, select the expression matrix (generated using LSTrAP). Using a | ||
normalized (TPM or RPKM) matrix is strongly recommended ! | ||
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Furthermore two additional files need to be provided, one that links the | ||
run identifiers to specific conditions. This tab delimited file should | ||
be structured as indicated below, a one-line header (which is ignored) | ||
and two columns, the first with the sample ID and the second with a short | ||
description of the condition sampled. Samples with the same description | ||
will be treated as replicates ! Omitting the condition description will | ||
exclude the sample from the profiles. | ||
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``` | ||
SampleID ConditionDescription | ||
SRR068987 Endosperm | ||
SRR314813 Seedlings, 11 DAG | ||
SRR314814 | ||
SRR314815 Flowers (floral buds) | ||
SRR314816 | ||
... | ||
``` | ||
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For profile plots on the website most likely a custum order of conditions | ||
is preferred. (We usually order tissues from bottom to top) A file to | ||
specify this needs to be provided, conditions need to be stated in the | ||
orther they should appear in the plot. | ||
Furthermore a color for that condition in the plot needs to be added in | ||
rgba() format. See the example below. | ||
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``` | ||
Roots (apex), 7 DAG rgba(153, 51, 0, 0.5) | ||
Roots (differentation zone), 4 DAP rgba(153, 51, 0, 0.5) | ||
Roots (elongation zone), 4 DAP rgba(153, 51, 0, 0.5) | ||
Roots (meristematic zone), 4 DAP rgba(153, 51, 0, 0.5) | ||
Roots (QC cells), 6 DAS rgba(153, 51, 0, 0.5) | ||
Roots (stele cells), 7 DAS rgba(153, 51, 0, 0.5) | ||
Roots (tip) rgba(153, 51, 0, 0.5) | ||
Leaves (rosette), 21 DAG rgba(0, 153, 51, 0.5) | ||
Leaves (rosette), 29 DAG rgba(0, 153, 51, 0.5) | ||
... | ||
``` | ||
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If all files are selected click 'Add Expression Profiles' to upload the | ||
data and add everything to the database. | ||
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