7
votes
How do I efficiently perform a metagenome screen of “all” species?
As you point out, BLAST is not going to work well for reads from a typical RNASeq experiment.
From an RNASeq dataset, you might be able to get away with just a ribosomal screen using Silva, because ...
- 13.8k
7
votes
How to use Python to count k-mers?
I wrote a command-line k-mer counter called kmer-counter that will output results in a form that your Python script can consume: https://github.com/alexpreynolds/...
- 3,125
5
votes
Accepted
How to use Python to count k-mers?
If you want to count the number of unique k-mers that occur in your data set, you should use the unique-kmers.py script, which implements a HyperLogLog-based cardinality estimator.
If the number of ...
- 5,060
4
votes
How do I efficiently perform a metagenome screen of “all” species?
I think you're just looking for nr. It is absolutely not limited to prokaryotes, far from it. It is, according to the blurb on the NCBI's blast page:
The ...
- 9,662
4
votes
What's the scaling for HOMER metagenes?
Figured it out! For anyone who finds this thread in the future and is wondering what is going on: HOMER, when using the -size given option, normalizes the y-axis to the number of basepairs in the ...
- 101
4
votes
How do I efficiently perform a metagenome screen of “all” species?
One thing that hasn't been suggested so far would be phylogenetic placement. First option there would be placing against one larger tree roughly touching the domains you mentioned to get an overview. ...
- 111
4
votes
How do I efficiently perform a metagenome screen of “all” species?
You might want to look into minihashes as a more efficient way of clustering and comparing your sequences to known species and samples. In the mash paper the authors show how it would be feasible to ...
- 967
4
votes
Accepted
4
votes
Accepted
Extract sequence context of high-degree nodes in assembly graphs
I don't have a recommendation of specific tools, but we might be able to use the GFA file itself to get at this with just bash commands.
For these examples I use the linked GFA here, which is a ...
- 3,804
4
votes
How do I measure the proportion of different bacteria in a sample from a high-throughput sequencer?
If you've got a fast solid state drive with >200G of space, then kraken2 + bracken works really well on both long and short reads (e.g. Nanopore, Illumina). It's also specifically designed for ...
- 13.8k
3
votes
NCBI SRA database sample: control vs test
You can download the information using this link:
I did it in R:
...
- 1,678
3
votes
Accepted
Transform traditional blast output to `--outfmt 6`
Biopython has a parser for Blast's default text format (which I assume is the one you have). Just use the blast-text string to specify this format in ...
- 967
3
votes
Transform traditional blast output to `--outfmt 6`
The blast_formatter command allows you to produce output in any of the formats supported by BLAST. However, it requires that you run ...
- 5,060
3
votes
Accidental mapping of eukaryotic reads in a metagenomic dataset
You can use centrifuge with the NT library to profile the taxonomy, and then remove the reads from eukaryotes.
- 59
3
votes
Accidental mapping of eukaryotic reads in a metagenomic dataset
If you know the eukaryotic contaminant present you could use bbsplit.sh from BBMap suite to split/bin reads first using a reference for that contaminant (into one ...
- 59
3
votes
Software for taxonomic assignment?
If you like blast, maybe you can try DIAMOND - the authors claim it's much faster than BLAST.
- 196
3
votes
Extract sequence context of high-degree nodes in assembly graphs
Edit: I've written a short blog post describing my use of nodeSeqs.sh to investigate the functions of sequences at the nodes of short-read assembly graphs for a set of human oral microbiome samples.
...
- 613
3
votes
Minimap2 detects too many 16S sequences in metagenome
16S is a very conserved sequence, which is why it's used for targeted phylogenetic analysis; it makes it easy to amplify and analyse. Unfortunately that conservation is an issue with minimap2, which ...
- 13.8k
3
votes
Accepted
How to make pan-core genome curve through command line on linux?
The command create_pan_genome_plots from Roary can create a plot like this:
- 387
3
votes
detecting DNA of possible infection on cell line
I've had some success looking for contamination in Illumina and Nanopore reads using kraken2.
Kraken2 has a phylogenetic-based approach, which in the first instance tries to find the most specific ...
- 13.8k
2
votes
Pooling data in metagenome assembly
With metagenomes, there's several strategies and no one-size-fits-all.
You can spend a lot of time playing with the data to try to get the "best assembly." But it depends on what you're trying to ...
- 106
2
votes
Accepted
Viral Metagenomics
The specifics will depend on the experimental setup and data, but a few general comments...
1) Metagenomic data is often large volume and high in microbial diversity but low and uneven in coverage; ...
- 3,928
2
votes
Accepted
Is there a way to assemble contigs starting from a specific sequence?
This is one of the primary use cases for which spacegraphcats (preprint and code) was designed. The "neighborhood queries" discussed in the paper sound particularly relevant. I don't have any personal ...
- 5,060
2
votes
Accepted
Software for microbial profiling from 16S rRNA gene sequence
I tried dada2 and is not bad (if you know R).
QIIME2 is also an option.
Many other are available, the choice might also depend on your sample and your exact question.
For functional profiling you ...
- 196
2
votes
Accepted
Comparing gene abundances between metagenomes
I would avoid using assemblies to answer this question, as there's no guarantee that you will be able to assemble your genes of interest; you can however estimate their abundance even if they are ...
- 3,804
2
votes
How identifiable are human omics data and how to mitigate their identifying features?
This is one of the major problems with genomic information in todays research. This was highlighted some years ago with the police using publicly available genomic data bases to idenitfy unkown murder ...
- 876
2
votes
Accepted
Reads count in metagenomics
I think that there is a good argument for using technologies adapted from RNA-seq for metagenomic abundance quantification. The tool that I am most familiar with (that I use for this purpose) is ...
- 3,804
2
votes
Accepted
Demovir produces an empty output
maybe you can open the demovir.R file with Rstudio and execute line by line to check where the failure is. - @zorbax
Thanks for your suggestion. I eventually got a correct final output already.
The ...
- 41
2
votes
Co-occurrence networks in Metagenomics studies
To answer your question more formally. I strongly believe you are treating the mice with something that will shift its microbial flora. Clearly the experiment is not described.
Thus what I simply feel ...
M__♦
- 11.9k
2
votes
Minimap2 detects too many 16S sequences in metagenome
You are taking reads as the reference. Given the similarity of 16S, you will find hits for most 16S in other species. You have to filter out poor hits. This is the faster way if you don't have many ...
- 6,485
Only top scored, non community-wiki answers of a minimum length are eligible