BOL: Related items

Variant Calling Resequencing-Based Genome Inference

Abhi — Wed, 31 Jul 2024 02:02:24 -0500

Variant Calling - Resequencing-Based Genome Inference

Erik Garrison
University of Tennessee Health Science Center
Workshop on Genomics - Český Krumlov
January 12, 2024

https://evomics.org/wp-content/uploads/2024/01/Variant-calling-Workshop-on-Genomics-2024-Cesky-Krumlov.pdf

Address of the bookmark: https://evomics.org/wp-content/uploads/2024/01/Variant-calling-Workshop-on-Genomics-2024-Cesky-Krumlov.pdf

Learning Python Programming - a bioinformatician perspective !

Rahul Nayak — Mon, 14 May 2018 16:33:03 -0500

Python Programming is a general purpose programming language that is open source, flexible, powerful and easy to use. One of the most important features of python is its rich set of utilities and libraries for data processing and analytics tasks. In the current era of big biological data, python and biopython is getting more popularity due to its easy-to-use features which supports big data processing.

In this tutorial series article, I will explore features and packages of python which are widely used in the big data, NGS, and bioinformatics. I will also walk through a real biological example which shows NGS data processing with the help of python packages and programming.

Python has a couple of points to recommend it to biologists and scientists specifically:

It's widely used in the scientific community
It has a couple of very well designed libraries for doing complex scientific computing (although we won't encounter them in this book)
It lend itself well to being integrated with other, existing tools
It has features which make it easy to manipulate strings of characters (for example, strings of DNA bases and protein amino acid residues, which we as biologists are particularly fond of)

In general, following are some of the important features of python which makes it a perfect fit for rapid application development.

Python is interpreted language so the program does not need to be compiled. Interpreter parses the program code and generates the output.
Python is dynamically typed, so the variables types are defined automatically.
Python is strongly typed. So the developers need to cast the type manually.
Less code and more use makes it more acceptable.
Python is portable, extendable and scalable.

There are two major Python versions, Python 2 and Python 3. Python 2 and 3 are quite different. This tutorial uses Python 3, because it more semantically correct and supports newer features.

I will post tutorial on daily basis on this page. Check the sub-pages on right side.

Interactive Bioinformatics Resources !

Jit — Thu, 12 Aug 2021 00:09:00 -0500

Learn how to use bioinformatics tools right from your browser.
Everything runs in a sandbox, so you can experiment all you want.

More at sandbox.bio

Address of the bookmark: http://sandbox.bio

Snakemake Tutorials !

Rahul Nayak — Mon, 09 May 2022 05:20:41 -0500

A lesson introducing the Snakemake workflow system for bioinformatics analysis.

Prerequisites

This is an intermediate lesson and assumes learners have already done some bioinformatics:

Familiarity with the BASH command shell, including concepts like pipes, variables and loops.

Knowledge of bioinformatics fundamentals like the FASTQ file format and transcriptome sequencing, in order to understand the example workflow.

No previous knowledge of Snakemake or workflow systems is required.

https://carpentries-incubator.github.io/snakemake-novice-bioinformatics/index.html

Address of the bookmark: https://carpentries-incubator.github.io/snakemake-novice-bioinformatics/aio/index.html

UnCoVar: Workflow for Transparent and Robust Virus Variant Calling, Genome Reconstruction and Lineage Assignment

BioStar — Mon, 05 Aug 2024 23:01:29 -0500

UnCoVar: Workflow for Transparent and Robust Virus Variant Calling, Genome Reconstruction and Lineage Assignment

Using state of the art tools, easily extended for other viruses
Tool and database updates for critical components via Conda
Built using modern design patterns with Conda and Snakemake
Extensible and easy to customize
Submission Ready Genomes
Customizable reporting with comprehensive visualization

https://ikim-essen.github.io/uncovar/

Github https://github.com/IKIM-Essen/uncovar

Address of the bookmark: https://ikim-essen.github.io/uncovar/

PHYMMBL

Jit — Mon, 10 Oct 2016 08:56:34 -0500

Metagenomics sequencing projects collect samples of DNA from uncharacterized environments that may contain hundreds or even thousands of species. One of the main challenges in analyzing a metagenome is phylogenetic classification of raw sequence reads into groups representing the same or similar species. Such classification is a useful prerequisite for genome assembly and for analysis of the biological diversity present in a sample. The newest sequencing technologies have simultaneously made metagenomics easier, by making the sequencing process faster, and more difficult, by producing shorter read lengths than previous technologies. Methods for classifying sequences as short as 100 base pairs (bp) have until now been relatively inaccurate, requiring metagenomics projects to use older, long-read technologies. Phymm, a new classification approach for metagenomics data which uses interpolated Markov models (IMMs) to taxonomically classify DNA sequences, can accurately classify reads as short as 100 bp. Its accuracy for short reads represents a significant leap forward over previous composition-based classification methods. PhymmBL (rhymes with "thimble"), the hybrid classifier included in this distribution which combines analysis from both Phymm and BLAST, produces even higher accuracy.

Address of the bookmark: http://www.cbcb.umd.edu/software/phymm/

A Beginner's Guide to Using Kraken for Taxonomic Classification

Neel — Fri, 13 Dec 2024 11:29:03 -0600

Kraken is a popular bioinformatics tool designed for fast and accurate taxonomic classification of metagenomic sequences. Its efficiency and precision make it a go-to resource for analyzing microbial communities, including bacteria, viruses, archaea, and fungi. Whether you're new to bioinformatics or experienced in the field, Kraken is an indispensable tool for taxonomic analysis.

In this blog, we’ll walk through the basics of Kraken, from installation to running an analysis, and highlight its key features and applications.

What is Kraken?

Kraken is a sequence classification tool that assigns taxonomic labels to DNA sequences using exact k-mer matching. It uses a reference database of genomes, dividing sequences into k-mers and identifying matches in a computationally efficient way.

Key Features of Kraken

Speed: Kraken processes data much faster than alignment-based methods.
Accuracy: It uses a precise k-mer matching algorithm for high-resolution taxonomic assignments.
Scalability: It can handle large metagenomic datasets.
Custom Databases: You can build and use custom databases tailored to your research needs.

Installing Kraken

System Requirements
- A Unix-based operating system (Linux/macOS).
- Sufficient computational resources for database building (RAM and disk space).
Installation Steps
- Clone the Kraken repository from GitHub:
  
  git clone https://github.com/DerrickWood/kraken.git cd kraken
- Compile the Kraken binaries:
  
  make
- Add Kraken to your PATH for easy access:
  
  export PATH=$PATH:/path/to/kraken

Preparing a Database

Kraken requires a database of reference genomes. You can use a pre-built database or create a custom one.

Downloading a Pre-built Database
Kraken offers pre-built databases, such as the MiniKraken database, which is lightweight and suitable for smaller datasets. Download it using:

kraken-build --download-library minikraken
Building a Custom Database
To include specific genomes, download FASTA files and build the database:

kraken-build --download-library bacteria --threads 4 --db my_database kraken-build --build --db my_database

This process may take considerable time and resources, depending on the size of the database.

Running Kraken

Once the database is ready, you can classify sequences.

Basic Usage
Use the following command to classify sequences:

kraken --db my_database --threads 4 --fastq-input input_sequences.fastq --output kraken_output.txt

Key options:
- --db: Specifies the database.
- --threads: Number of threads for parallel processing.
- --fastq-input: Indicates input file format (FASTQ/FASTA).
Interpreting Results
Kraken generates an output file with columns for sequence IDs, taxonomic classifications, and the confidence score.

Visualizing Kraken Results

Kraken results can be visualized using tools like Krona or converted to human-readable reports using kraken-report.

Generate a Report

kraken-report --db my_database kraken_output.txt > kraken_report.txt
Krona Visualization
Install Krona and convert Kraken output for visualization:

cut -f2,3 kraken_output.txt | ktImportTaxonomy -o krona_output.html

Open the HTML file in your browser to interactively explore the taxonomic classifications.

Advanced Usage

Confidence Thresholds
Adjust the confidence threshold for classification using the --confidence option. Higher values reduce false positives but may miss some true positives:

kraken --db my_database --confidence 0.1 --fastq-input input.fastq
Paired-End Reads
For paired-end sequencing data, use:

kraken --db my_database --paired reads_1.fastq reads_2.fastq
Customizing K-mers
Kraken allows you to set custom k-mer lengths during database building for specific applications.

Applications of Kraken

Microbial Ecology: Characterizing microbial communities in soil, water, and the human microbiome.
Pathogen Detection: Identifying pathogens in clinical samples.
Fungal Research: Analyzing fungal diversity in metagenomic datasets.
Environmental Monitoring: Tracking microbial populations in diverse habitats.

Conclusion

Kraken is a versatile and efficient tool for taxonomic classification in metagenomics. Its speed, accuracy, and flexibility make it a favorite among bioinformaticians. By following this guide, you can set up and use Kraken to unlock insights into microbial and fungal communities, paving the way for discoveries in ecology, medicine, and biotechnology.

CLARK: Fast, accurate and versatile sequence classification system

Jit — Sat, 15 Feb 2020 01:49:01 -0600

CLARK, a method based on a supervised sequence classification using discriminative k-mers. Considering two distinct specific classification problems (see the article for details), namely (1) the taxonomic classification of metagenomic reads to known bacterial genomes, and (2) the assignment of BAC clones and transcript to chromosome arms/centromeres (in the absence of a finished assembly for the reference genome), CLARK outperforms in classification speed and precision the best state-of-the-art methods.

http://clark.cs.ucr.edu/Spaced/

Address of the bookmark: http://clark.cs.ucr.edu/Spaced/

Understanding DUMP files from NCBI Taxonomy database !

Shruti Paniwala — Fri, 15 Jul 2022 04:29:05 -0500

*.dmp files are bcp-like dump from GenBank taxonomy database

General information.

Field terminator is "\t|\t"

Row terminator is "\t|\n"

nodes.dmp file consists of taxonomy nodes. The description for each node includes the following

fields:

tax_id -- node id in GenBank taxonomy database

parent tax_id -- parent node id in GenBank taxonomy database

rank -- rank of this node (superkingdom, kingdom, ...)

embl code -- locus-name prefix; not unique

division id -- see division.dmp file

inherited div flag (1 or 0) -- 1 if node inherits division from parent

genetic code id -- see gencode.dmp file

inherited GC flag (1 or 0) -- 1 if node inherits genetic code from parent

mitochondrial genetic code id -- see gencode.dmp file

inherited MGC flag (1 or 0) -- 1 if node inherits mitochondrial gencode from parent

GenBank hidden flag (1 or 0) -- 1 if name is suppressed in GenBank entry lineage

hidden subtree root flag (1 or 0) -- 1 if this subtree has no sequence data yet

comments -- free-text comments and citations

Taxonomy names file (names.dmp):

tax_id -- the id of node associated with this name

name_txt -- name itself

unique name -- the unique variant of this name if name not unique

name class -- (synonym, common name, ...)

Divisions file (division.dmp):

division id -- taxonomy database division id

division cde -- GenBank division code (three characters)

division name -- e.g. BCT, PLN, VRT, MAM, PRI...

comments

Genetic codes file (gencode.dmp):

genetic code id -- GenBank genetic code id

abbreviation -- genetic code name abbreviation

name -- genetic code name

cde -- translation table for this genetic code

starts -- start codons for this genetic code

Deleted nodes file (delnodes.dmp):

tax_id -- deleted node id

Merged nodes file (merged.dmp):

old_tax_id -- id of nodes which has been merged

new_tax_id -- id of nodes which is result of merging

Citations file (citations.dmp):

cit_id -- the unique id of citation

cit_key -- citation key

pubmed_id -- unique id in PubMed database (0 if not in PubMed)

medline_id -- unique id in MedLine database (0 if not in MedLine)

url -- URL associated with citation

text -- any text (usually article name and authors).

-- The following characters are escaped in this text by a backslash:

-- newline (appear as "\n"),

-- tab character ("\t"),

-- double quotes ('\"'),

-- backslash character ("\\").

taxid_list -- list of node ids separated by a single space

Dengue Lineages !

LEGE — Fri, 16 Aug 2024 04:40:14 -0500

Our dengue virus lineage system splits up the current genotypes into major and minor lineages to provide additional spatiotemporal resolution and a common language to discuss important genomic diversity. A full description of the lineage system can be found here.

https://dengue-lineages.org/

Address of the bookmark: https://dengue-lineages.org/