Tag: NCBI Prokaryotic Genome Annotation Pipeline (PGAP)

New version of PGAP available now!

We are happy to announce that a new version of PGAP is available. This version will annotate 20 to 25% more genes with symbols (e.g. recA) on the assembled genomes of key species, compared to previous versions.

You will observe an increase in symbols when you annotate the genomes of Escherichia coli, Campylobacter jejuni and a few other species. As several users have requested, this feature will facilitate the comparison of gene content across multiple genomes. It is permitted by the addition of a new workflow to PGAP for identifying orthologs between the reference genomes of Escherichia coli str. K-12 substr. MG1655, Bacillus subtilis subsp. subtilis str. 168, Campylobacter jejuni subsp. jejuni NCTC 11168, Mycobacterium tuberculosis H37Rv, and Acinetobacter pittii PHEA-2 and genomes in the same species being annotated. Symbols of reference genes with defined function are propagated to their orthologs in the genome annotated with PGAP.

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Assemble and annotate your prokaryotic genomes with RAPT

Do you need an easy way to analyze a bacterium you just isolated? The latest version of NCBI’s Read assembly and Annotation Pipeline Tool (RAPT) is a pilot web service for the assembly and gene annotation of public or private Illumina genomic reads sequenced from bacterial or archaeal isolates.

We’ll be giving a webinar on webRAPT on May 19 where you can learn more, but you can test it out now.

Get started with the click of a button

RAPT is simple to use.

1. If you’re working with NIH’s Sequence Read Archive (SRA) and have an SRA accession, enter it in the first box below (Figure 1a) or upload a file of sequencing reads in the second box (Figure 1b).

screenshot of webRAPT submission; enter SRA accession number on the left or upload files on the right
Figure 1. 1a, on the left. Enter an SRA run accession (starting with SRR, DRR or ERR) in the text box on the left if you wish to assemble reads that are already public and press submit. If you are providing a read set that is not in SRA, use the box on the right, shown in 1b. Enter the organism name (genus only or genus species known to NCBI Taxonomy) in the “Organism” field. Click “One file” if all reads for the run are in a single file. This file can contain single-end reads or paired-end reads with reads of a pair adjacent to each other in the file (interleaved). Upload the sequencing reads using the “Choose file” button. Click “Two files” to provide forward and reverse reads from a paired-end library in two separate files. Upload the forward and reverse files using the “Choose Forward Reads File” and “Choose Reverse Reads File” buttons. Then press submit.

 

 

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May 19 Webinar: Using the new web RAPT service to assemble and annotate prokaryotic genomes

May 19 Webinar: Using the new web RAPT service to assemble and annotate prokaryotic genomes

Join us on May 19, 2021 at 12PM eastern time to learn how to use the new  RAPT pilot service to assemble and annotate public or private Illumina genomic reads sequenced from bacterial or archaeal isolates at the click of a button. RAPT consists of two major components, the genome assembler SKESA and the Prokaryotic Genome Annotation Pipeline (PGAP), and produces an annotated genome of quality comparable to RefSeq in a couple of hours.

  • Date and time: Wed, May 19, 2021 12:00 PM – 12:45 PM EDT
  • Register

After registering, you will receive a confirmation email with information about attending the webinar. A few days after the live presentation, you can view the recording on the NCBI webinars playlist on the NLM YouTube channel. You can learn about future webinars on the Webinars and Courses page.

NCBI at CSHL Biology of Genomes, May 11 – 14, 2021

NCBI at CSHL Biology of Genomes, May 11 – 14, 2021

NCBI staff will be presenting virtual posters at the Cold Spring Harbor Laboratory  Biology of Genomes Meeting, May 11 -14, 2021. The posters will cover the following topics: 1) a cloud-ready suite of tools (PGAP, RAPT , and SKESA) for assembling and annotating prokaryotic genomes,  2) Datasets — a new set of services for downloading genome assemblies and annotations, and 3) updates on NCBI RefSeq eukaryotic genome annotation, and the Genome Data Viewer (GDV). Read more below for the full abstracts.

The virtual poster gallery opens Tuesday, May 11 at 9:00 a.m. with dedicated time for poster viewing and discussion at 1:00 to 2:00 p.m. through Slack each day. The poster gallery will be open for entire the conference and remain available for six weeks afterwards.  Continue reading “NCBI at CSHL Biology of Genomes, May 11 – 14, 2021”

Improvements to NCBI Assembly

NCBI’s genome Assembly has a number of significant improvements!

Assembly records now have a link to Primer-BLAST making it easy to design primers in the context of a specific eukaryote genome assembly.  Figure 1 shows the Assembly page for the Genome Reference Consortium Mouse Build 39 (GRCm39) with the link to Primer-BLAST.

Figure 1. The Assembly page for the mouse reference genome (GCF_000001635.27). Showing the new Run Primer-BLAST link, which loads the assembly as a database in the Primer-BLAST search (bottom) and the new expandable note sections, Genome-Annotation-Data in this case. 
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New release of the Read Assembly and Annotation Pipeline Tool (RAPT), now 2X faster!

There is a new release of the Read assembly and Annotation Pipeline Tool (RAPT) available from our GitHub site. RAPT is a one-step application for the genome assembly and gene annotation of archaeal and bacterial isolates that can run on your local computer or the Google Cloud Platform (GCP). With this new release, jobs will run twice as fast as with the December release. For example, we have assembled and annotated a Salmonella enterica genome in under an hour on a 16-CPU machine with the new release.
We have also added several new features based on your feedback including:

  1. The –stop-on-errors flag that will stop the process if there evidence from the average nucleotide identity check that there is sample mix-up or contamination by other bacteria.
  2. The ability to accept forward and reverse reads of paired-end runs in separate files. These can be compressed (gzip) files.

Finally, thanks to all who came to our webinar in December and provided their comments! For these who couldn’t join us, you can now view the recording on our YouTube channel.

Contact us at prokaryote-tools@ncbi.nlm.nih.gov with any question and to let us know if you would like to become a beta-tester for RAPT.

NCBI hidden Markov models (HMM) release 4.0 now available!

Release 4.0 of the NCBI hidden Markov models (HMM) used by the Prokaryotic Genome Annotation Pipeline (PGAP) is now available from our FTP site. You can search this collection against your favorite prokaryotic proteins to identify their function using the HMMER sequence analysis package.

This release contains 17,443 models, including 94 new models since the last release. We have also updated names and added EC numbers and  gene symbols to over 100 models. You can search and view the details of these HMMs in the newly deployed Protein Family Model collection that also includes conserved domain architectures and BlastRules  and allows you to find all RefSeq proteins named by these profiles. See our recent post for more details.

The Protein Family Model resource is now available!

The new Protein Family Model resource  (Figure 1) provides a way for you to search across the evidence used by the NCBI annotation pipelines to name and classify proteins. You can find protein families by gene symbol, protein function, and many other terms. You have access to related proteins in the family and publications describing members. Protein Family Models includes protein profile hidden Markov models (HMMs) and BlastRules for prokaryotes, and conserved domain architectures for prokaryotes and eukaryotes. The HMMs in the collection include Pfam models, TIGRFAMs as well as models developed at NCBI either de novo, or from NCBI protein clusters.  Each of the BlastRules (PMCID: 5753331) consists of one or more model proteins of known biological function with BLAST identity and coverage cutoffs.  The conserved domain architectures are based on BLAST-compatible Position Specific Score Matrices  (PSSMs) that constitute the NCBI Conserved Domain database.Figure 1. Protein Family Model resource pages. Top panel.  Home page. Middle  panel, selected results summaries from a fielded search for the DnaK gene product (DnaK[Gene Symbol]). Bottom panel, a portion of an HMM record for DnaK derived from NCBI Protein Clusters (NF009946). The record also includes PubMed citations and HMMER analyses showing the RefSeq proteins named by this method.

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RefSeq Release 202 is public

RefSeq release 202 is accessible online, via FTP and through NCBI’s Entrez programming utilities, E-utilities.

This full release incorporates genomic, transcript, and protein data available as of September 8, 2020, and contains 255,571,455 records, including 186,755,483 proteins, 33,077,068 RNAs, and sequences from 104,969  organisms. The release is provided in several directories as a complete dataset and also as divided by logical groupings.

Updated human genome Annotation Release 109.20200815
Updated Annotation Release 109.2020815 is an update of NCBI Homo sapiens Annotation Release 109. The annotation report is available here.

The annotation products are available in the sequence databases and on the FTP site.

This update includes around 15,000 updated RefSeq transcripts revised to use CAGE and polyA data to define 5′ and 3′ ends, and match the reference GRCh38 sequence.

Coronavirus host gene regulatory elements now annotated by RefSeq Functional Elements
The RefSeq Functional Elements project at NCBI has prioritized curation of experimentally validated regulatory elements for human host genes associated with SARS-CoV-2 entry into cells. The annotations include several enhancers, promoters, cis-regulatory elements and protein binding sites, among other feature types. We annotated 236 regulatory features for 27 distinct biological regions, including regulatory elements for the ABO, ACE2, ANPEP, CD209, CLEC4G, CLEC4M, CTSL, DPP4, and TMPRSS2 genes. More information can be found here.

New eukaryotic genome annotations
This release includes new annotations generated by NCBI’s eukaryotic genome annotation pipeline for 27 species, including:

  • maize annotation release 103, based on the new assembly Zm-B73-REFERENCE-NAM-5.0 (GCF_902167145.1)
  • marmoset annotation release 105, based on the new assembly Callithrix_jacchus_cj1700_1.1 (GCF_009663435.1)
  • Chinese hamster annotation release 104, based on the assembly CriGri_1.0 (GCF_000223135.1) and the new assembly CriGri-PICRH-1.0 (GCF_003668045.3)
  • Asian giant hornet annotation release 100, based on the new assembly V.mandarinia_Nanaimo_p1.0 (GCF_014083535.2)
  • Florida lancelet annotation release 100, based on the new assembly Bfl_VNyyK (GCF_000003815.2)
  • Anopheles stephensi annotation release 100, based on the new assembly UCI_ANSTEP_V1.0 (GCF_013141755.1)

Updated and improved collection of RefSeq representative genome assemblies now available
The collection of representative genome assemblies for Bacteria and Archaea contains 11,727 prokaryotic assemblies to represent their respective species. More information can be found here.

Updated protein family models used by PGAP available for download
Release 3.0 of the NCBI protein family models used by the Prokaryotic Genome Annotation Pipeline (PGAP) is now available.

This release contains 17,350 models: 12,864 HMMs built at NCBI (111 more than in release 2.0) and 4,486 TIGRFAM HMMs. In addition, since release 2.0, we have assigned product names to over 2,000 Pfam HMMs, bringing the total to 6,698 Pfam HMMs with names that can be transferred by PGAP to the annotated proteins they hit. More information can be found here.

Future change: Mouse Reference Assembly Update
RefSeq annotation of the new mouse GRCm39 assembly is in progress, and is expected to be included in the next release.

Updated and improved collection of RefSeq representative genome assemblies now available

We have updated the collection of representative genome assemblies for Bacteria and Archaea. As announced in April, this set is now recalculated three times a year. We selected a total of 11,727 prokaryotic assemblies to represent their respective species among the 192,000 assemblies in RefSeq. Six hundred and thirty-five species were included in the collection for the first time, while 395 organisms from undefined species (such as Bacillus bacterium) were removed. We were able to choose a higher-quality representative than in the previous set for 18% of Bacterial and Archaeal species due to improvements in the logic of the selection that is now based on the assembly length, number of pseudo CDSs called in the PGAP annotation, number of scaffolds, whether Gene IDs are available in the Gene database for the assembly that is currently representative, and type strain status. You can see the exact criteria in order of importance on the Prokaryotic RefSeq Genomes page. Now that the new selection process is in place, we expect future updates to have fewer changes. We will replace a representative only if the assembly has changed RefSeq status or if a substantially better assembly becomes available.

We have updated the database on the Microbial Nucleotide BLAST page as well as the basic nucleotide BLAST RefSeq Representative Genome Database, to reflect these changes.

You can download the reference and representative set from the Assembly resource. If you are interested in the annotation on these genomes, you can limit searches to proteins annotated on representative genomes by adding “refseq_select[filter]” to any query in the Protein database. For example, you can find all proteins annotated on representative genomes in the genus Klebsiella by using the query: “Klebsiella[organism] AND refseq_select[filter]“.  A BLAST database of proteins annotated on representative genomes will be coming soon. Stay tuned!