web-based nextclade issue when using another reference

[yl315504]

Hello,

I was trying to use a mouse covid reference, called MA10.
https://www.ncbi.nlm.nih.gov/nuccore/1898953378

I uploaded the fasta file as the new reference. Should I use fasta file as the reference?

I got the following error. Could you please help?

Thanks,

Error message: Error: When initializing Nextclade runner: When parsing reference tree Auspice JSON v2: When parsing Auspice Tree JSON contents: When parsing JSON: expected value at line 1 column 1

Nextclade version 2.14.1 (commit: 85e00e8, branch: release)

Memory available: 3586 MBytes

User agent: Mozilla/5.0 (Windows NT 10.0; Win64; x64) AppleWebKit/537.36 (KHTML, like Gecko) Chrome/119.0.0.0 Safari/537.36

Browser details: {"browser":{"name":"Chrome","version":"119.0.0.0"},"os":{"name":"Windows","version":"NT 10.0","versionName":"10"},"platform":{"type":"desktop"},"engine":{"name":"Blink"}}

Call stack:

Error: When initializing Nextclade runner: When parsing reference tree Auspice JSON v2: When parsing Auspice Tree JSON contents: When parsing JSON: expected value at line 1 column 1
at M (https://clades.nextstrain.org/_next/static/chunks/68.daad1778960b5734.js:1:15774)
at https://clades.nextstrain.org/_next/static/wasm/cf835b4f9fbcf56c.wasm:wasm-function[1547]:0x1e3d5c
at new a (https://clades.nextstrain.org/_next/static/chunks/68.daad1778960b5734.js:1:9424)
at https://clades.nextstrain.org/_next/static/chunks/68.daad1778960b5734.js:1:24056
at u (https://clades.nextstrain.org/_next/static/chunks/444.170c49d0571b2ba1.js:1:28086)
at Generator._invoke (https://clades.nextstrain.org/_next/static/chunks/444.170c49d0571b2ba1.js:1:29376)
at a. [as next] (https://clades.nextstrain.org/_next/static/chunks/444.170c49d0571b2ba1.js:1:28489)
at p (https://clades.nextstrain.org/_next/static/chunks/68.daad1778960b5734.js:1:21841)
at d (https://clades.nextstrain.org/_next/static/chunks/68.daad1778960b5734.js:1:22038)
at https://clades.nextstrain.org/_next/static/chunks/68.daad1778960b5734.js:1:22097

[corneliusroemer]

Hi @yl315504! Unfortunately you can't just swap out a reference when the dataset contains a tree that is based on a particular reference. If the genemap is different that might also cause issues - unfortunately this isn't as simple as plugging in your own reference.

What's the reason you'd like to use this particular reference? There might be another way to achieve your goal.

If you just want to align against that reference, you can use nextalign CLI.

[ivan-aksamentov]

Dear @yl315504,

On the screenshot, I see you have a red circle with "1" inside of it, near "Customize dataset files". This means you've provided some custom dataset files under that section (the section can be opened and closed).

From the error, saying that Nextclade failed to parse Auspice JSON, I hypothesize that you've provided a reference tree file there which is not a correctly formatted Auspice JSON tree file. This means that you either need to provide a correctly formatted reference tree file or to remove it (in which case the default one will be used, from the SC2 dataset).

The part you've encircled with a red line is the list of files which Nextclade will be analyzing (we call them "query sequences").

Note that each dataset is tailored towards a particular pathogen strain. In general, you cannot just swap one component and expect everything to work as usual. For example, as a general rule, reference sequence must be a root of the reference tree (there are workarounds, but they are quite advanced). So only slight customizations are possible. I don't believe analyzing viruses from different host organisms are possible with our human SC2 dataset, unless the strains are very close to Wuhan strain or other pandemic strains. You can always create your own whole dataset though.

So all this looks very confusing to me. I invite you to read Nextclade documentation (the "Docs" link on the top panel) It's important to understand how Nextclade works and how to configure it before using it, and especially before using its advanced features. It's not the best documentation in the world, but it might worth to give it a shot. If you still have questions after reading documentation, we'll try to answer.

Please explain better what you are trying to achieve and we'll try to help.

[yl315504]

Thanks for your reply. Our goal is to explore if our sequences have new mutations against the mouse covid MA-10. Is there a way to do this using the web-based nextclade? Thanks, Yan Liu UT Southwestern Medical Center

…

On Wed, Dec 20, 2023 at 3:11 PM Cornelius Roemer ***@***.***> wrote: Hi @yl315504 <https://github.com/yl315504>! Unfortunately you can't just swap out a reference when the dataset contains a tree that is based on a particular reference. If the genemap is different that might also cause issues - unfortunately this isn't as simple as plugging in your own reference. What's the reason you'd like to use this particular reference? There might be another way to achieve your goal. If you just want to align against that reference, you can use nextalign CLI. — Reply to this email directly, view it on GitHub <#1364 (comment)>, or unsubscribe <https://github.com/notifications/unsubscribe-auth/AGCZ2OD6OJ5HVJDKXHYL4RTYKNH7TAVCNFSM6AAAAABA5MU7I2VHI2DSMVQWIX3LMV43OSLTON2WKQ3PNVWWK3TUHMYTQNRVGE2DGNBZG4> . You are receiving this because you were mentioned.Message ID: ***@***.***>

-- Thanks, *Yan Liu*

[corneliusroemer]

Thanks for explaining your goal! It's possible right now but not easy - but we might be able to implement something to make this easier, I'll break it out into a feature request.

Right now, I don't think we have the option to start with a "blank" dataset and add your own files onto it.

Currently, it's a bit tricky to do this. One way to achieve this would be to create your own dataset: https://docs.nextstrain.org/projects/nextclade/en/stable/user/datasets.html

You might be best off running Nextclade v3 CLI like this:

nextclade3 run --input-ref mouse-seq.fasta --output-tsv results.tsv input.fasta

This is much easier than creating a dataset. You could look at the output tsv in Excel and see whether there are any mutations.

I'll think about how to make things easier for this use case. Thanks for reaching out in any case, it's always useful to know what end users struggle with and what their use cases are!

[yl315504]

Thanks! I will use Nextclade v3 CLI. nextclade3 run --input-ref mouse-seq.fasta --output-tsv results.tsv input.fasta Yan Liu

…

On Wed, Dec 20, 2023 at 3:37 PM Cornelius Roemer ***@***.***> wrote: Thanks for explaining your goal! It's possible right now but not easy - but we might be able to implement something to make this easier, I'll break it out into a feature request. Right now, I don't think we have the option to start with a "blank" dataset and add your own files onto it. Currently, it's a bit tricky to do this. One way to achieve this would be to create your own dataset: https://docs.nextstrain.org/projects/nextclade/en/stable/user/datasets.html You might be best off running Nextclade v3 CLI like this: nextclade3 run --input-ref mouse-seq.fasta --output-tsv results.tsv input.fasta This is much easier than creating a dataset. You could look at the output tsv in Excel and see whether there are any mutations. I'll think about how to make things easier for this use case. Thanks for reaching out in any case, it's always useful to know what end users struggle with and what their use cases are! — Reply to this email directly, view it on GitHub <#1364 (comment)>, or unsubscribe <https://github.com/notifications/unsubscribe-auth/AGCZ2OA7LGL4QZN6OD37OHDYKNLARAVCNFSM6AAAAABA5MU7I2VHI2DSMVQWIX3LMV43OSLTON2WKQ3PNVWWK3TUHMYTQNRVGE3TMOJQGE> . You are receiving this because you were mentioned.Message ID: ***@***.***>

-- Thanks, *Yan Liu*

[corneliusroemer]

@yl315504 Nextclade v3 is not yet conda-installable, so you need to download the binary from here: https://github.com/nextstrain/nextclade/releases/tag/3.0.0-alpha.1

However, as an early christmas present for you (I hope) I just implemented a basic dataset that you can drop your own reference into (the thing I suggested earlier we could possibly do).

This is very experimental and might not work for very long, but you can give it a try here:
https://nextclade-git-cds-error-nextstrain.vercel.app/?dataset-server=gh:@scratch@&dataset-name=nextstrain/scratch/reference-only

You need to "customize" the dataset as shown in the video, but otherwise it should work now (in contrast to your earlier attempts!):

Would love to know if this works and if it does what you need it to do.

[yl315504]

It works! This is the best Christmas gift! Yan Liu

…

On Wed, Dec 20, 2023 at 5:07 PM Cornelius Roemer ***@***.***> wrote: @yl315504 <https://github.com/yl315504> Nextclade v3 is not yet conda-installable, so you need to download the binary from here: https://github.com/nextstrain/nextclade/releases/tag/3.0.0-alpha.1 However, as an early christmas present for you (I hope) I just implemented a basic dataset that you can drop your own reference into (the thing I suggested earlier we could possibly do). This is very experimental and might not work for very long, but you can give it a try here: ***@***.***@&dataset-name=nextstrain/scratch/reference-only You need to "customize" the dataset as shown in the video, but otherwise it should work now (in contrast to your earlier attempts!): 2023-12-21.00.03.16.gif (view on web) <https://github.com/nextstrain/nextclade/assets/25161793/2a4da92e-b9bc-4386-af27-b0bb4e0463c3> Would love to know if this works and if it does what you need it to do. — Reply to this email directly, view it on GitHub <#1364 (comment)>, or unsubscribe <https://github.com/notifications/unsubscribe-auth/AGCZ2OCXPDTTAA7LCZBMPLLYKNVS3AVCNFSM6AAAAABA5MU7I2VHI2DSMVQWIX3LMV43OSLTON2WKQ3PNVWWK3TUHMYTQNRVGI2TQMJTGU> . You are receiving this because you were mentioned.Message ID: ***@***.***>

-- Thanks, *Yan Liu*

[corneliusroemer]

Excellent! Right now the URL you need to use is quite ugly, we might make this a normal dataset so it's easier to select in the future.

If you have other ideas/use cases/requests, let us know and we can see whether it's possible.