BOL: Related items

Chemical Elements of Bioinformatics

Rahul Agarwal — Tue, 03 Sep 2013 16:35:39 -0500

You must be familiar with periodic table and colour pattern, but this time you are going to amaze by new elements table by Eagle genomics. Just check it out and have fun :)

http://elements.eaglegenomics.com/

BLAST options, setting and defaults

Jit — Mon, 10 Dec 2018 08:29:37 -0600

BLAST stands for Basic Local Alignment Search Tool and was developed by Altschul et al. (1990) and significantly improved by Altschul et al. (1997). It is a very fast search algorithm that is used to separately search protein or DNA databases. BLAST is best used for sequence similarity searching, rather than for motif searching. For searches using a query sequence of fewer than twenty residues, PatMatch is the best choice. Another sequence alignment tool that may yield different results from BLAST, and may be useful for motif searching, is FASTA. To search nonplant datasets, try NCGR BLAST or NCBI BLAST.

A fairly complete on-line guide to BLAST searching can be found at the NCBI BLAST Help Manual. For a theoretical overview of BLAST, see the NCBI BLAST Course. Additional information can be found in the BLAST 2.0 Release Notes

	BLASTN	BLASTP	BLASTX	TBLASTN	TBLASTX	PSIBLAST
Gap opening penalty: cost to open a gap [integer]	default = 5	default = 11 limited values are supported	default = 11 limited values are supported	default = 11 limited values are supported	default = 11 limited values are supported	default = 5
Gap extension penalty: cost to extend a gap [integer]	default = 2	default = 1 a 0 in this field means to use the default	default = 1 a 0 in this field means to use the default	default = 1 a 0 in this field means to use the default	default = 1 a 0 in this field means to use the default	default = 2
Nucleic match: reward for a match in the BLAST portion of run [integer]	default = 1	n/a	n/a	n/a	n/a	default = 1
Nucleic mismatch: penalty for a mismatch in the blast portion of run [integer]	default = -3	n/a	n/a	n/a	n/a	default = -3
Expectation value: (E) [real]	default = 10.0	default = 10.0	default = 10.0	default = 10.0	default = 10.0	default = 10.0
Word size: the size of the initial word that must be matched between the database and the query sequence	default = 11	default = 3	default = 3	default = 3	default = 3	default = 11
Max scores: Number of one-line descriptions (V) [Integer]	default = 25	default = 25	default = 25	default = 25	default = 25	default = 25
Max alignments: number of alignments to show (B) [integer]	default = 15	default = 15	default = 15	default = 15	default = 15	default = 15
Query filter: filter applied to the query sequence	default = DUST	default = SEG	default = SEG	default = SEG	default = SEG	default = DUST
Query genetic code: genetic code to be used in BLASTX translation of the query	n/a	n/a	default = universal	default = universal	default = universal	n/a
Matrix: substitution matrix to be used for amino acid comparisons	no default	default = blosum62	default = blosum62	default = blosum62	default = blosum62	no default

Supported and Suggested Values for Gap Open and Extension in BLASTP, BLASTX, TBLASTN, and TBLASTX

Gaps Open	Gap Extension
10	1
10	2
11	1
8	2
9	2

Address of the bookmark: https://www.arabidopsis.org/Blast/BLASToptions.jsp

NCBIBLAST+ 2.14.1 now available

Neel — Wed, 30 Aug 2023 02:36:13 -0500

#NCBIBLAST+ 2.14.1 now available with improved documentation, faster and more reliable database downloads, and some bug fixes.

Check out the changes they made.

They added the cleanup-blastdb-volumes.py script to remove unused BLAST database volumes. Read the documentation here.

They also switched the protocol from ftp to https to access BLAST databases for increased performance and reliability when downloading data from the NCBI with the update_blastdb.pl script.

And fixed a few bugs related to downloading data from the NCBI, and mt_mode crashing blastn and blastx.

Check out the release notes.

Download BLAST+ 2.14.1

Questions or comments? Please write the BLAST help desk.

More info and download: https://blast.ncbi.nlm.nih.gov/doc/blast-news/2023-BLAST-News.html

BLAST+ 5: Key Updates and Enhancements for Modern Bioinformatics

LEGE — Sat, 07 Dec 2024 22:37:48 -0600

The BLAST+ 5 (Basic Local Alignment Search Tool) update has introduced several key enhancements aimed at improving performance, user experience, and compatibility with evolving genomic data standards. Here are the major updates:

Database Enhancements:
- The BLAST databases have shifted fully to the version 5 (v5) format, which integrates built-in taxonomy information. This allows for more detailed and efficient sequence annotation and analysis.
- Protein databases in v5 are now accession-based, supporting a broader range of sequences, including those from high-throughput projects and the Pathogen Detection Project. These databases also accommodate structural proteins with multi-character chain identifiers.
Performance Improvements:
- Adaptive Composition-Based Statistics (CBS) is available as an experimental feature, enhancing the detection of novel results in protein-protein comparisons.
- Updated algorithms improve the stability of search results, especially when fewer hits are requested than the default output.
Compatibility:
- Support for the older v4 databases has been discontinued. The v5 format is now the default for all BLAST database updates, ensuring alignment with current standards in bioinformatics.
User-Friendly Changes:
- Naming conventions for databases have been simplified to enhance clarity and ease of use. For example, database names no longer include version tags like "_v5".
Future-Proofing:
- BLAST+ 5 aligns with current and upcoming data requirements, ensuring that researchers have access to the most comprehensive and modern resources for sequence alignment.

These updates reflect NCBI's commitment to maintaining BLAST as a leading tool for sequence analysis. For detailed release notes and additional guidance, refer to NCBI Insights here

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.

Demo 4: Using BLAST/BLAT in Ensembl

Tue, 10 Sep 2013 11:54:03 -0500

We demonstrate the BLAST/BLAT tool in Ensembl. Search for a sequence in Ensembl, and identify hits to the genome, or to genes, with this tool.

Genome Workbench 2.10.7

Gudiya Pal — Fri, 01 Jul 2016 12:09:59 -0500

Genome Workbench 2.10.7 is here! New features include added support for local custom BLAST databases and improvements to Tree View.

For the full list of features, improvements and fixes, see the release notes:https://ncbi.nlm.nih.gov/tools/gbench/releasenotes

New Features

BLAST Tool: added support for local custom BLAST databases
Graphical Sequence View: added log scaling option for graph tracks
Generic Table View: new tutorial added

Bug Fixes and Improvements

Project Tree View: Genomic Collections/Assemblies now show accessions, not just names
Tree View: layout updated to better accommodate nodes of different sizes
Table Import Dialog (MacOS): fixed issue with table visibility
Fixed bug where different molecules IDs in GenBank could resolve to the same sequence
Graphical Sequence View: fixed issue where sequence track was not shown for some sequences
Graphical Sequence View: fixed protein coloration methods
Graphical Sequence View: improved rendering of Markers to better indicate boundaries and produce higher quality PDF images
Create Gene Model tool: fixed scenario when gene model tool failed with local sequences
Search View: ORF Finder – fixed incorrect protein lengths
Fixed bug with not opening project file (.gbp) on a click
Fixed issues in GVF import
Fixed BLAST Search tool against NCBI databases not working
Fixed tblastn (protein BLAST) not working in standalone mode
Fixed GTF export failure

DIAMOND

Jit — Thu, 27 Apr 2017 04:21:54 -0500

DIAMOND is a sequence aligner for protein and translated DNA searches and functions as a drop-in replacement for the NCBI BLAST software tools. It is suitable for protein-protein search as well as DNA-protein search on short reads and longer sequences including contigs and assemblies, providing a speedup of BLAST ranging up to x20,000.

More at file:///home/urbe/Downloads/diamond_manual.pdf

http://www.nature.com/nmeth/journal/v12/n1/full/nmeth.3176.html

Address of the bookmark: https://github.com/bbuchfink/diamond

BLAST nr version 5 database, (nr_v5)

Jit — Fri, 23 Aug 2019 11:35:35 -0500

NCBI have made changes the nr version 5 database, (nr_v5), to facilitate better search results and improved performance by reducing the number of redundant titles in the nr_v5 database used by webBLAST, which is also available for BLAST+ users.

The changes in nr preserve the taxonomic diversity of the entries in the database while reducing the number of titles for identical sequences. GenPept accessions are still accessible via www.ncbi.nlm.nih.gov/protein/$GENBANK_ACCESSION or the IPG website https://www.ncbi.nlm.nih.gov/ipg/.

The "Identical Proteins" link in the alignments section of the webBLAST results takes you to a full list of all accessions associated with a sequence.

For BLAST+ users downloading nr_v5: the database is now approximately 50% smaller, resulting in faster downloads and BLAST searches, and smaller disk space requirements. The database is downloadable at: ftp://ftp.ncbi.nlm.nih.gov/blast/db/v5/

For BLAST+ there is a cleanup script to help you manage the transition to this smaller database. The script removes unused database volumes: ftp://ftp.ncbi.nlm.nih.gov/blast/temp/cleanup-blastdb-volumes.py

Here are the new rules on how we keep titles in nr_v5:

1. We keep all refseq, swissprot, pir and PDB titles.

2. We keep any GenPept titles with a TAXID that has not already been seen in the record.

3. We keep at least five GenPept titles regardless of whether the TAXIDS have been seen before or not in this record.

Primer BLAST !

BioStar — Tue, 28 Apr 2020 00:28:49 -0500

BLAST team added a new feature (Max 3' match), shown in Figure 1, to Primer-BLAST that limits the length of 3' exon matches when designing exon-exon spanning primers. This makes it less likely that primers specifically designed to amplify transcripts will also amplify genomic DNA contamination in expression assays. See the NCBI Insights post (https://go.usa.gov/xvUT4) for more details.

If you have any questions or concerns, please contact blast-help@ncbi.nlm.nih.gov