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TRAMP: a software package for generating transposon maps.

TRAMP is a software package for generating transposon maps that are used for DNA sequencing. The package provides a variety of automated tools that can always be overridden by the user. The central part of the package is its selection algorithm that finds the most robust map with the smallest number of inserts. TRAMP has been in daily use by the sequencing team at LBL since it was introduced in the Spring of 1994. It is applicable to any sequencing project utilizing the directed strategy.

Algorithms↗

GEOMETRY: a software package for nucleotide sequence analysis using statistical geometry in sequence space.

GEOMETRY is a software package for the analysis of nucleotide sequences using the method of statistical geometry in sequence space. The package consists of programs performing estimation of the average geometry of sequence quartets, analysis of positional variability and computer simulation of parallel and tree-like sequence divergence with user-defined parameters. It provides an independent tool for evaluation of the reliability of conventional phylogenetic trees and calibration of the time of sequence divergence. GEOMETRY may be of interest for all scientists engaged in the study of molecular phylogeny. The package is available by anonymous FTP from ftp.bionet.nsk.su, directory /incoming/molevol/geom.exe, and will be available from EMBL file server (URL: http://@www.ebi.ac.uk)

Algorithms↗

Mathematica packages for simulation of experimental genetics.

UNLABELLED: This note describes add-on packages for the Mathematica software system (Wolfram 1996) which allow simulation and analysis of both Mendelian and complex genetic traits in experimental crosses of plants or animals. AVAILABILITY: The add-on packages are freely available at http://www.mathsource.com/cgi-bin/msitem?0209-30 4. SUPPLEMENTARY INFORMATION: A tutorial notebook file is included with the packages at the mathsource site.

Animals↗

CMAP: contig mapping and analysis package, a relational database for chromosome reconstruction.

In the contig mapping and analysis package, CMAP, we provide a foundation for reverse genetics by organizing information about DNA fragments obtained from an organism's genome into a physical map. The user can store information about a particular segment of DNA. This information can be both descriptive, such as any genes contained in a particular DNA fragment, or experimental, such as hybridization profiles or restriction digest patterns for comparison with other fragments. The package can then be instructed to update the physical map or provide information on a DNA fragment within the map, such as its location. The user interface is designed to minimize the learning curve associated with database usage, while eliminating the possibility of entering data outside the ranges of fields through error-checking protocols. Queries are currently accomplished by the use of dynamic SQL (structured query language), which gives the user the ability to build queries based on any combination of the attributes contained within the database without requiring that all possible queries be permanently programmed within the query software. In order to eliminate the need for knowledge of SQL, an interface was designed to allow users to build queries by menu choices. Thus, CMAP is a software package supporting a database for both the production and storage of a physical map as well as being the first step toward the production of a physical mapping workstation.

Algorithms↗

PCAP: probe choice and analysis package--a set of programs to aid in choosing synthetic oligomers for contig mapping.

In the program, PCAP, we provide a methodology for choosing synthetic oligonucleotide probes to be used in contig mapping experiments. The package serves the purpose of presenting a series of short oligonucleotides (8-12mers) that are chosen based on constraints with respect to frequency of occurrence within a particular genome and the G+C content of the oligonucleotides. The four programs contained within the package: (i) convert GenBank files to a format useable by the package; (ii) calculate trinucleotide and tetranucleotide frequencies in available sequence data on a particular species; (iii) present the user with upper and lower bounds on the frequencies of hybridization sites for oligonucleotide probes of length 8-12, (iv) allow the user to place constraints on site frequency and G+C content and provides a list of short probe sequences that fit these criteria. These sequences can then be synthetically produced and used in hybridization experiments to carry out contig mapping.

Base Sequence↗

SpectroPipeR-a streamlining post Spectronaut® DIA-MS data analysis R package.

SUMMARY: Proteome studies frequently encounter challenges in down-stream data analysis due to limited bioinformatics resources, rapid data generation, and variations in analytical methods. To address these issues, we developed SpectroPipeR, an R package designed to streamline data analysis tasks and provide a comprehensive, standardized pipeline for Spectronaut® DIA-MS data. This novel package automates various analytical processes, including XIC plots, ID rate summary, normalization, batch and covariate adjustment, relative protein quantification, multivariate analysis, and statistical analysis, while generating interactive HTML reports for e.g. ELN systems. AVAILABILITY AND IMPLEMENTATION: The SpectroPipeR package (manual: https://stemicha.github.io/SpectroPipeR/) was written in R and is freely available on GitHub (https://github.com/stemicha/SpectroPipeR).

Software↗

moiraine: an R package to construct reproducible pipelines for the application and comparison of multi-omics integration methods.

MOTIVATION: In the past decades, many statistical methods for integrating multi-omics data have been developed. They have been implemented into software tools, which differ widely in their programming choices, such as the format required for data input, or the format of the generated integration results. This lack of standards renders cumbersome and time-intensive the application and comparison of different integration tools to the same multi-omics dataset. RESULTS: We have developed the moiraine R package for constructing reproducible multi-omics integration pipelines, which enables users to apply one or more statistical methods for multi-omics integration to their own multi-omics dataset. moiraine facilitates the preprocessing of the omics datasets and automates their formatting for the integration step. It simplifies the interpretation and evaluation of the integration results through the construction of visualizations in which metadata about samples and features can easily be included. Crucially, it enables the comparison of results obtained with different integration tools, allowing users to assess the robustness of their results. AVAILABILITY AND IMPLEMENTATION: The moiraine R package is publicly available at https://github.com/Plant-Food-Research-Open/moiraine; an archival snapshot of the package is available on Zenodo at https://doi.org/10.5281/zenodo.17172718. A detailed tutorial is available at https://plant-food-research-open.github.io/moiraine-manual/.

Software↗

phylobar: an R package for multiresolution compositional barplots in omics studies.

SUMMARY: Stacked barplots, though widely used in microbiome studies, can obscure important patterns in microbiome data. They omit rare taxa and can mask shifts that emerge at finer taxonomic levels. To address this issue, we introduce phylobar, an R package that interactively links stacked barplots with overview phylogenetic or taxonomic hierarchies. The interface allows users to collapse or expand subtrees, paint color palettes interactively, and search for specific taxa. This allows comparison across taxonomic resolutions that are hidden in static overviews. phylobar works with any omics data with hierarchical organization, including cell type hierarchies, as we demonstrate in a case study of immune cell composition in COVID-19 patients. AVAILABILITY AND IMPLEMENTATION: phylobar is available as an R package on GitHub. It uses the htmlwidgets library to link interactive D3 visualizations with R. The interactive plots can be embedded within R Markdown or Quarto notebooks, and views can be exported as vector graphics files. The package is open source and documented at https://mkdiro-O.github.io/phylobar.

Software↗

CAP: conformation angles package-displaying the conformation angles of side chains in proteins.

SUMMARY: A graphics package has been developed to display all side chain conformation angles of the user selected residue in a given protein structure. The proposed package is incorporated with all the protein structures (solved using X-ray diffraction and NMR spectroscopy) available in the Protein Data Bank. The package displays the multiple conformations adopted by a single amino acid residue whose structure is solved and refined at very high resolution. In addition, it shows the percentage distribution of the side chain conformation angles in different rotameric states. AVAILABILITY: http://144.16.71.146/cap/

Amino Acids↗

matchprobes: a Bioconductor package for the sequence-matching of microarray probe elements.

UNLABELLED: The nucleotide sequences of the probes on a microarray can be used for a variety of purposes in the analysis of microarray experiments. We describe software and a paradigm for the creation of data packages for curating, distributing and working with probe sequence data in a uniform, across-types-of-microarrays manner. While the implementation is specific to the Bioconductor project, the ideas and general strategies are more general and could be easily adopted by other projects. AVAILABILITY: The R package matchprobes is available under LGPL at http://www.bioconductor.org SUPPLEMENTARY INFORMATION: The package contains documentation in the form of a vignette and manual pages.

Algorithms↗

OntologyTraverser: an R package for GO analysis.

UNLABELLED: Gene Ontology (GO) annotations have become a major tool for analysis of genome-scale experiments. We have created OntologyTraverser--an R package for GO analysis of gene lists. Our system is a major advance over previous work because (1) the system can be installed as an R package, (2) the system uses Java to instantiate the GO structure and the SJava system to integrate R and Java and (3) the system is also deployed as a publicly available web tool. AVAILABILITY: Our software is academically available through http://franklin.imgen.bcm.tmc.edu/OntologyTraverser/. Both the R package and the web tool are accessible. CONTACT: cashaw@bcm.tmc.edu

Algorithms↗

Bio3d: an R package for the comparative analysis of protein structures.

UNLABELLED: An automated procedure for the analysis of homologous protein structures has been developed. The method facilitates the characterization of internal conformational differences and inter-conformer relationships and provides a framework for the analysis of protein structural evolution. The method is implemented in bio3d, an R package for the exploratory analysis of structure and sequence data. AVAILABILITY: The bio3d package is distributed with full source code as a platform-independent R package under a GPL2 license from: http://mccammon.ucsd.edu/~bgrant/bio3d/

Amino Acid Sequence↗

A novel method for generating nested deletions using the in vitro bacteriophage T3 DNA packaging system.

To sequence a DNA segment inserted into a cosmid vector under the directed sequencing strategy, we established a simple and rapid method for generating nested deletions which uses the in vitro packaging system of bacteriophage T3 DNA. The principle is based on the previous finding that this system can translocate any linear double-stranded DNA up to 40 kb into the phage capsid in a time-dependent manner and the encapsulated DNA becomes DNase-resistant. For this purpose, we constructed a cosmid vector that carries two different antibiotic selection markers at both sides of the multiple cloning site, and after insertion of a DNA segment, the clone was linearized by lambda-terminase at the cos site. After the packaging reaction in vitro followed by DNase treatment, the encapsulated DNA was introduced into Escherichia coli cells to give clones with unidirectional deletions by differential antibiotic selection. Restriction and sequence analyses of deletion clones demonstrated that an ordered set of clones with nested deletions, ranging from less than 1 kb to 25 kb, was created from either the end of the DNA segment. Thus, nested deletion clones that cover the entire region of a approximately 40-kb cosmid insert can be obtained by a single packaging reaction, and its restriction map can be simultaneously obtained.

Bacteriophage T3↗

An improved cosmid vector for the nested deletion method using the bacteriophage T3 DNA packaging system.

We constructed a new cosmid vector suitable for the previously developed nested deletion method which used the in vitro DNA packaging system of bacteriophage T3. The first step of this method is linearization of a cosmid clone to be packaged, and we previously introduced cleavage at the cos site using lambda-Terminase, but optimization of the reaction conditions was required for complete digestion because of its instability. In the newly constructed vector, pAT5, the sites of 4 different restriction enzymes, Sse8387I, Asc I, Fse I and Pme I, each of which recognizes an 8-bp sequence (8-base cutter) were introduced in the vicinity of the cos site. In addition, the species of restriction sites for cloning were increased to broaden its application. The cosmid clone constructed by this new vector could be linearized at one of the 8-base cutter sites which are assumed to rarely occur in the genome, and followed by in vitro packaging, nested deletion clones were successfully prepared.

Bacteriophage T3↗

The acceptability to primary care staff of a multidisciplinary training package on acute back pain guidelines.

BACKGROUND: Implementing clinical guidelines is more likely to be successful when the whole practice team is committed to the process. Practices from the MRC General Practice Research Framework in two distinct geographical centres in the UK (West Yorkshire and Greater Manchester) participated in the feasibility study for the UK Back pain Exercise And Manipulation (UK BEAM) trial. Practice teams were randomized to continue with their usual care for back pain patients, or to be trained in managing back pain in line with national guidelines. Those randomized to the intervention arm of the trial were invited to attend training, delivered by either a generic trainer or a back pain expert. OBJECTIVES: Our aims were to assess the general acceptability of the training package to staff, to assess the acceptability of the multidisciplinary approach and to determine if a generic primary care educator could deliver the training as effectively as a clinical back pain expert. METHODS: All staff (clinical and non-clinical) from intervention practices were invited to attend multidisciplinary training sessions on the active management of back pain. Practice staff in West Yorkshire were trained by a generic primary care educator and practice staff in Greater Manchester were trained by a clinical back pain expert. The content of sessions was standardized for both trainers and included didactic and interactive components and small group, case study discussions. Detailed notes were taken of observations made of participants during sessions, and evaluation forms were completed by all those who attended. RESULTS: The majority of participants found the training useful and said that the session had lived up to their expectations. Most found that the session was well planned and that they had sufficient opportunity to participate in learning. The training package was well received by clinical staff, but was less acceptable to non-clinical staff. GPs dominated the small group work discussions. No differences were found between the preferences of participants for the two different trainers. CONCLUSION: The training package was appropriate for clinical staff, but did not always meet the needs of non-clinical staff and may require modification for this group. A generic educator can successfully lead multidisciplinary educational sessions addressing clinical issues.

Acute Disease↗

Genetic evidence that recognition of cosQ, the signal for termination of phage lambda DNA packaging, depends on the extent of head filling.

Packaging a phage lambda chromosome involves cutting the chromosome from a concatemer and translocating the DNA into a prohead. The cutting site, cos, consists of three subsites: cosN, the nicking site; cosB, a site required for packaging initiation; and cosQ a site required for termination of packaging. cosB contains three binding sites (R sequences) for gpNu1, the small subunit of terminase. Because cosQ has sequence identity to the R sequences, it has been proposed that cosQ is also recognized by gpNu1. Suppressors of cosB mutations were unable to suppress a cosQ point mutation. Suppressors of a cosQ mutation (cosQ1) were isolated and found to be of three sorts, the first affecting a base pair in cosQ. The second type of cosQ suppression involved increasing the length of the phage chromosome to a length near to the maximum capacity of the head shell. A third class of suppressors were missense mutations in gene B, which encodes the portal protein of the virion. It is speculated that increasing DNA length and altering the portal protein may reduce the rate of translocation, thereby increasing the efficiency of recognition of the mutant cosQ. None of the cosQ suppressors was able to suppress cosB mutations. Because cosQ and cosB mutations are suppressed by very different types of suppressors, it is concluded that cosQ and the R sequences of cosB are recognized by different DNA-binding determinants.

Bacteriophage lambda↗

Differential chromatin packaging of genomic imprinted regions between expressed and non-expressed alleles.

Chromosomal regions subject to genomic imprinting comprise a functional domain exhibiting parental-specific expression of genes and hence may take a unique chromatin structure. Here we have examined the chromatin packaging state of allelic sites in the Zfp127/Snrpn locus on mouse chromosome 7 and in the Igf2r locus on mouse chromosome 17 with an assay consisting of chromatin fractionation and allele-specific detection. The results showed that non-transcribed alleles of Igf2r are packaged more compactly than transcribed alleles in F(1) hybrid mice of both types of cross between C57BL/6 and MSM strains, whereas a non-imprinted gene, Sod-2, in the vicinity of Igf2r does not show such a difference. This indicates a close correlation between imprinting and the differential packaging of chromatin. On the other hand, the Zfp127/Snrpn locus showed such an allele-specific fractionation pattern only in F(1) hybrid mice of a cross but not in those of the reciprocal cross. Analysis of the congenic mice produced for this locus did not provide any difference. These results suggest that chromatin of imprinted domains in different compaction levels is affected by distinct blueprints in homologous chromosomes that are heritable through the germ line.

Alleles↗

A convenient and adaptable package of DNA sequence analysis programs for microcomputers.

We describe a package of DNA data handling and analysis programs designed for microcomputers. The package is convenient for immediate use by persons with little or no computer experience, and has been optimized by trial in our group for a year. By typing a single command, the user enters a system which asks questions or gives instructions in English. The system will enter, alter, and manage sequence files or a restriction enzyme library. It generates the reverse complement, translates, calculates codon usage, finds restriction sites, finds homologies with various degrees of mismatch, and graphs amino acid composition or base frequencies. A number of options for data handling and printing can be used to produce figures for publication. The package will be available in ANSI Standard FORTRAN for use with virtually any FORTRAN compiler.

Amino Acid Sequence↗