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Publish all current staging branch changes #442

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02e55c2
Adding in some style with css
cansavvy Oct 7, 2020
9e70344
Use css magic
cansavvy Oct 7, 2020
a7aad2f
Try making the navbar blue
cansavvy Oct 7, 2020
3927752
Add survey link
cansavvy Oct 15, 2020
8d7b8e2
Make font smaller
cansavvy Oct 15, 2020
d3acd2f
Merge branch 'master' into cansavvy/refinebio-fashion
cansavvy Oct 15, 2020
b0ced0f
Need a comma
cansavvy Oct 15, 2020
a7a17cd
Merge branch 'master' into cansavvy/refinebio-fashion
cansavvy Oct 16, 2020
de9726b
Change to normalizePath
cansavvy Oct 19, 2020
6928056
normalizepath separate step references.bib
cansavvy Oct 19, 2020
4c07c28
Move references.bib to component folder
cansavvy Oct 19, 2020
fa14538
Update github actions to reflect staging branch (#311)
cansavvy Oct 21, 2020
4ab1d80
Add google analytics to renderings (#314)
cansavvy Oct 21, 2020
de3dccc
Merge branch 'master' into staging
cansavvy Oct 21, 2020
1f2546c
Only push if we are in master.
jashapiro Oct 21, 2020
7d7ecc1
Add test target
jashapiro Oct 21, 2020
b96130e
test staging workflow with this branch
jashapiro Oct 21, 2020
a0640eb
back to latest tag
jashapiro Oct 21, 2020
8afc962
Try separate push step
jashapiro Oct 21, 2020
6a38574
change tags to test push
jashapiro Oct 21, 2020
f71bccc
Revert "change tags to test push"
jashapiro Oct 21, 2020
9de0042
Remove this branch from triggers
jashapiro Oct 21, 2020
28e72ef
Push staging, retag and push master
jashapiro Oct 22, 2020
a660e89
Made ccs modifications, added logo file
dvenprasad Oct 22, 2020
a24971a
Merge remote-tracking branch 'origin/master' into cansavvy/refinebio-…
cansavvy Oct 22, 2020
c6158e8
Resolve render-notebooks.R conflict
cansavvy Oct 22, 2020
145fd98
Remove testing html from file diff
cansavvy Oct 22, 2020
43b9ea7
uncommented mobile nav
dvenprasad Oct 22, 2020
b055f63
Merge pull request #321 from AlexsLemonade/jashapiro/dont-push-stagin…
jashapiro Oct 22, 2020
ca85ee0
Merge branch 'staging' into cansavvy/refinebio-fashion
cansavvy Oct 22, 2020
b1644c2
Update scripts/render-notebooks.R
jashapiro Oct 22, 2020
d87fce8
Merge pull request #325 from AlexsLemonade/cansavvy/refinebio-fashion
jashapiro Oct 22, 2020
6e57f15
Add some issue templates (#319)
cansavvy Oct 22, 2020
e56e60d
Update diagrams showing how microarray/RNA-seq work (#326)
cbethell Oct 22, 2020
36b81e8
Adding basic footer (#307)
cansavvy Oct 23, 2020
9a0474c
Updating CONTRIBUTING.md with instructions about staging -> master se…
cansavvy Oct 27, 2020
718de40
New PR templates to help with new process (#334)
cansavvy Oct 27, 2020
93c6c65
Try a "main PR" strategy with links to the real PR templates (#337)
cansavvy Oct 29, 2020
033f0cd
Add contributing diagrams (#333)
cansavvy Oct 29, 2020
70e58bb
Make the "Other" PR template the default (#341)
cansavvy Oct 29, 2020
01771e8
Add timeline reminder to issue template (#342)
cansavvy Oct 29, 2020
489cc04
Pr 1 of 2: Add Microarray Pathway Analysis - GSEA example (#345)
cbethell Nov 6, 2020
05a68f7
Pr 2 of 2: Add Microarray Pathway Analysis - GSEA example (#347)
cbethell Nov 16, 2020
936025b
Delete intro Rmd and renumber (#355)
cansavvy Nov 16, 2020
0f6a663
Add words about Draft and Refined PRs to CONTRIBUTING.md (#361)
cansavvy Nov 19, 2020
6e8579f
WGCNA Part 1: Set up (#358)
cansavvy Nov 19, 2020
4d82d89
WGCNA Part 2: Running WGCNA (#360)
cansavvy Nov 20, 2020
114d6fe
Add pathway analysis intro paragraph to microarray ORA (#356)
cansavvy Nov 20, 2020
ba7e18c
Fix WGCNA installation (#366)
jashapiro Nov 20, 2020
d6cfc8a
Pr 1 of 2: Add Microarray Pathway Analysis - GSVA example (#359)
cbethell Nov 20, 2020
2fc9de2
WGCNA Part 3: DE and heatmaps (#363)
cansavvy Nov 23, 2020
2658508
WGCNA Part 4: Warn about Outliers (#364)
cansavvy Nov 24, 2020
ae9aea8
Microarray ORA Restructure Instruction (#377)
cansavvy Nov 25, 2020
3a2f92e
WGCNA Part 5: switch dataset (#379)
cansavvy Nov 25, 2020
2dd6738
Change pdf -> png and rereun (#382)
cansavvy Nov 30, 2020
7c7555a
Pr 2 of 2: Add Microarray Pathway Analysis - GSVA example (#362)
cbethell Nov 30, 2020
ede4bc3
Remove getting started zip file (#392)
cansavvy Dec 1, 2020
458068f
ORA RNA-seq: Part 1 - The Set Up (#394)
cansavvy Dec 1, 2020
aace680
ORA RNA-seq: Part 2 - Run ORA and get results! (#395)
cansavvy Dec 2, 2020
9acefe5
Add message = FALSE to mute chatty blocks (#398)
jashapiro Dec 2, 2020
b2eb394
add GSVA package to Dockerfile (#401)
cbethell Dec 2, 2020
cc19310
Add rendering options via include.R (#402)
jashapiro Dec 2, 2020
4b8b557
GSVA for RNA-seq Part 1: Set up (#403)
cansavvy Dec 3, 2020
cb39d7c
Link citations in render (#407)
jashapiro Dec 3, 2020
9274aaf
Try different strategy for ortholog file download (#411)
cansavvy Dec 4, 2020
9601726
Editing/polish of microarray heatmap notebook (#409)
jashapiro Dec 4, 2020
47a62b4
Carry over common comment changes (#414)
jashapiro Dec 4, 2020
c2d911c
Use same download.file strategy for ortholog RNA-seq example (#413)
cansavvy Dec 4, 2020
94649c6
GSVA for RNA-seq: Part 2 -- GSVA and a heatmap (#404)
cansavvy Dec 5, 2020
beb8805
RNA-seq DGE dataset switch (#416)
cansavvy Dec 7, 2020
076ec33
add part 1 of RNA-seq GSEA example notebook (take 2) (#419)
cbethell Dec 8, 2020
f728c51
PCA polishing edits (#421)
jashapiro Dec 8, 2020
b676351
Try out a different download strategy for ORA (#418)
cansavvy Dec 8, 2020
4629244
Use download.file for the three other notebooks (#422)
cansavvy Dec 8, 2020
770b81b
PR 2 of 2: Add RNA-seq Pathway Analysis - GSEA example (take 2) (#420)
cbethell Dec 9, 2020
ac4a938
rerun Snakefile to fix html output (#424)
cbethell Dec 10, 2020
4b043ce
Umap polish (#423)
jashapiro Dec 10, 2020
f188c7c
Move filtering to before DESeq2 object creation (#425)
cansavvy Dec 11, 2020
5deefa9
Heatmap polish (#426)
jashapiro Dec 14, 2020
5b304a8
Polish differential exp microarray notebooks (#427)
jashapiro Dec 16, 2020
0af63a6
Minor Polish Diff Expr RNAseq (#429)
jashapiro Dec 16, 2020
94ee41a
Polish the microarray ORA notebook (#430)
jaclyn-taroni Dec 16, 2020
8ea03bc
Part 1: Add rownames.print = FALSE where its helpful (#431)
cansavvy Dec 16, 2020
fe4f9f7
Polish the RNA-seq ORA example (#432)
jaclyn-taroni Dec 16, 2020
2ea88fa
Add the rownames.print = FALSE and re-render (#433)
cansavvy Dec 16, 2020
0dc67a1
Polish Microarray GSEA example (#434)
jaclyn-taroni Dec 18, 2020
6f6f02b
Polish RNA-seq GSEA example (#437)
jaclyn-taroni Dec 18, 2020
11f0877
GHA: Slack us if Docker build or rendering fails (#438)
jaclyn-taroni Dec 20, 2020
a8ddcf4
Polish Ensembl Gene ID conversions (bonus reference updates!) (#435)
jashapiro Dec 21, 2020
a28c126
Polish microarray GSVA example (#440)
jaclyn-taroni Dec 21, 2020
b35d7a8
Polishing Ortholog notebooks (#436)
jashapiro Dec 21, 2020
4a54881
Polish the RNA-seq GSVA example (#441)
jaclyn-taroni Dec 21, 2020
4d9ba88
Merge remote-tracking branch 'origin/staging' into publish-all-changes
cansavvy Dec 21, 2020
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11 changes: 11 additions & 0 deletions .github/workflows/docker-build-push.yml
Original file line number Diff line number Diff line change
Expand Up @@ -91,3 +91,14 @@ jobs:
git add -A
git commit -m 'Render html and publish' || echo "No changes to commit"
git push origin gh-pages || echo "No changes to push"

# If we have a failure, Slack us
- name: Report failure to Slack
if: always()
uses: ravsamhq/[email protected]
with:
status: ${{ job.status }}
notify_when: 'failure'
env:
SLACK_WEBHOOK_URL: ${{ secrets.ACTION_MONITORING_SLACK }}
SLACK_MESSAGE: 'Build, Render, and Push failed'
11 changes: 11 additions & 0 deletions .github/workflows/docker-build.yml
Original file line number Diff line number Diff line change
Expand Up @@ -42,3 +42,14 @@ jobs:
tags: ccdl/refinebio-examples:latest
cache-from: type=local,src=/tmp/.buildx-cache
cache-to: type=local,dest=/tmp/.buildx-cache

# If we have a failure, Slack us
- name: Report failure to Slack
if: always()
uses: ravsamhq/[email protected]
with:
status: ${{ job.status }}
notify_when: 'failure'
env:
SLACK_WEBHOOK_URL: ${{ secrets.ACTION_MONITORING_SLACK }}
SLACK_MESSAGE: 'Build Docker failed'
1 change: 1 addition & 0 deletions .gitignore
Original file line number Diff line number Diff line change
Expand Up @@ -10,6 +10,7 @@ _site
*/plots/*
*/results/*
*/data/*
*/gene_sets/*

# markdown spellcheck
.spelling
Expand Down
916 changes: 875 additions & 41 deletions 01-getting-started/getting-started.html

Large diffs are not rendered by default.

30 changes: 17 additions & 13 deletions 02-microarray/00-intro-to-microarray.Rmd
Original file line number Diff line number Diff line change
Expand Up @@ -9,7 +9,7 @@ output:

Data analyses are generally not "one size fits all"; this is particularly true when with approaches used to analyze RNA-seq and microarray data.
The characteristics of the data produced by these two technologies can be quite different.
This tutorial has example analyses [organized by technology](../01-getting-started/getting-started.html#about-how-this-tutorial-book-is-structured) so you can follow examples that are more closely tailored to the nature of the data at hand.
This tutorial has example analyses [organized by technology](https://alexslemonade.github.io/refinebio-examples/01-getting-started/getting-started.html#about-how-this-tutorial-book-is-structured) so you can follow examples that are more closely tailored to the nature of the data at hand.

<!-- START doctoc generated TOC please keep comment here to allow auto update -->
<!-- DON'T EDIT THIS SECTION, INSTEAD RE-RUN doctoc TO UPDATE -->
Expand All @@ -28,15 +28,15 @@ This tutorial has example analyses [organized by technology](../01-getting-start

## Introduction to microarray technology

Microarrays measure gene expression using chips filled with oligonucleotide probes designed to hybridize to labeled RNA samples.
Microarrays measure gene expression using chips filled with oligonucleotide probes designed to hybridize to labeled RNA samples.
After hybridization, the microarrays are scanned, and the fluorescence intensity for each probe is measured.
The fluorescence intensity indicates the number of labeled fragments bound and therefore the relative quantity of the transcript the probe is designed for.

<img src="https://github.com/AlexsLemonade/refinebio-examples/raw/46f3d93471088218eda3104aa7a62bd90f6dfa0c/components/figures/microarray-overview.png" width=600>

[based on diagram from @microarray-video]

There are many different kinds of microarray platforms, which can be broadly separated into single-color and [two-color arrays](https://www.ebi.ac.uk/training/online/course/functional-genomics-ii-common-technologies-and-data-analysis-methods/microarrays).
There are many different kinds of microarray platforms, which can be broadly separated into single-color and [two-color arrays](https://www.ebi.ac.uk/training/online/course/functional-genomics-ii-common-technologies-and-data-analysis-methods/microarrays).
At this time, refine.bio only supports single-color arrays, so our examples and advice are generally from the perspective of using single-color array.
The diagram above shows an overview of the single-color array process which includes extracting the total RNA from a sample, labeling the RNA with fluorescent dye, hybridizing the labels, and scanning the fluorescent image to analyze the fluorescence intensity.

Expand All @@ -45,27 +45,31 @@ A longer list of specific arrays that are supported by refine.bio can be found [

As with all experimental methods, microarrays have strengths and limitations that you should consider in regards to your scientific questions.

### Microarray data **strengths**:
### Microarray data **strengths**:

- Microarray is generally less expensive than RNA-seq - you can afford more replicates and get higher statistical power [@Tarca2006].
- Microarray has generally had a faster turn-around than RNA-seq [@LCSciences2014].
- Microarrays historically were less expensive than RNA-seq allowing for more replicates and greater statistical power [@Tarca2006].
- Microarrays generally had a faster turn-around than RNA-seq [@LCSciences2014].

### Microarray data **limitations**:
As a result of these historical advantages, vast quantities of data have been generated worldwide using microarrays.
The microarray data compiled by refine.bio includes over 500,000 individual samples across over 25,000 experiments.
For many scientific questions, the best available gene expression data may be microarray based!

- If a transcript doesn't have a probe designed to it on a microarray, it won't be measured; standard microarrays can't be used for transcript discovery [@Mantione2014].
### Microarray data **limitations**:

- If a transcript doesn't have a probe designed to it on a microarray, it won't be measured; standard microarrays can't be used for transcript discovery [@Mantione2014].
- A chip's probe designs are only as up to date as the genome annotation at the time it was designed [@Mantione2014].
- As is true for all techniques that involve nucleotide hybridization (RNA-seq too); microarray probes come with some biases depending on their nucleotide sequence composition (like GC bias).

Refine.bio drops outdated probes based on [Brainarray’s annotation packages](http://brainarray.mbni.med.umich.edu/Brainarray/Database/CustomCDF/) and uses [SCAN](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3508193/pdf/nihms401888.pdf) normalization methods prior to your downloads to help address these probe nucleotide composition biases [@Dai2005; @Piccolo2012].
refine.bio drops outdated probes based on [Brainarray’s annotation packages](http://brainarray.mbni.med.umich.edu/Brainarray/Database/CustomCDF/) and uses [SCAN](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3508193/pdf/nihms401888.pdf) normalization methods prior to your downloads to help address these probe nucleotide composition biases [@Dai2005; @Piccolo2012].

## About quantile normalization

Microarray chips are generally experimentally processed in groups of chips - this can lead to [experimental batch effects](https://en.wikipedia.org/wiki/Batch_effect#:~:text=In%20molecular%20biology%2C%20a%20batch,of%20interest%20in%20an%20experiment).
To minimize this, all refine.bio microarray data downloads come [quantile-normalized](https://en.wikipedia.org/wiki/Quantile_normalization) which enables more confident comparisons of expression levels among experiments.
Different microarray chips are also a type of batch effect, but quantile normalization allows us to compare data from different chips that to a limited degree if we proceed with caution.
The use of different microarray chips is also a type of batch effect, but quantile normalization allows us to compare data from different chips to a limited degree, if we proceed with caution.
See the refine.bio docs for more about the microarray processing steps, including the [quantile normalization](http://docs.refine.bio/en/latest/main_text.html#quantile-normalization).

## More resources on microarray technology:
## More resources on microarray technology:

- [Getting started in gene expression microarray analysis](https://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1000543) [@Slonim2009].
- [Microarray and its applications](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3467903/) [@Govindarajan2012].
Expand All @@ -79,13 +83,13 @@ See the refine.bio docs for more about the microarray processing steps, includin

- A common and simple reason you may not see your gene of interest is that the microarray chip used in the experiment you are analyzing did not originally have probes designed to target that gene.

- Refine.bio uses [Brainarray packages](http://brainarray.mbni.med.umich.edu/Brainarray/Database/CustomCDF/) to annotate the microarray probe data for microarray platforms that have this available [@Dai2005].
- refine.bio uses [Brainarray packages](http://brainarray.mbni.med.umich.edu/Brainarray/Database/CustomCDF/) to annotate the microarray probe data for microarray platforms that have this available [@Dai2005].
This annotation identifies which probes map to which genes according to the updated transcriptome annotation (which likely changed since the microarray’s probes were first designed).
Some probes may have since become obsolete (they do not bind reliably to one location according to updated genome annotations), which may result in the gene they targeted being removed.
If your gene of interest was covered by the original probes of the microarray chip and the version of the Brainarray package used maintains that it is still accurate, your gene of interest will show up in the Gene column.
You can find your dataset’s microarray chip and Brainarray version information on the refine.bio dataset page and [by following these instructions](TODO: Put link to refine.bio docs FAQ when https://github.com/AlexsLemonade/refinebio-docs/issues/137 is addressed).

- One additional reason you may not see a gene of interest applies only if you are refine.bio's [aggregate by species](https://docs.refine.bio/en/latest/main_text.html#aggregations) option.
When data is aggregated across different platforms, only the genes common to both/all experiments aggregated will be kept.
When data is aggregated across different platforms, only the genes common to all experiments aggregated will be kept.

## References
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