Evidence map›Paper›PMID 33648451›Full record

ArticleBMC bioinformatics2021

barCoder: a tool to generate unique, orthogonal genetic tags for qPCR detection.

Casey B Bernhards, Matthew W Lux, Sarah E Katoski, Tyler D P Goralski, Alvin T Liem, Henry S Gibbons

Abstract read
In one paragraph

Article in BMC bioinformatics, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

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PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors.

Casey B Bernhards *U.S. Army Combat Capabilities Development Command Chemical Biological Center, Aberdeen Proving Ground, MD, 21010, USA.
Matthew W Lux *U.S. Army Combat Capabilities Development Command Chemical Biological Center, Aberdeen Proving Ground, MD, 21010, USA.
Sarah E KatoskiU.S. Army Combat Capabilities Development Command Chemical Biological Center, Aberdeen Proving Ground, MD, 21010, USA.
Tyler D P GoralskiU.S. Army Combat Capabilities Development Command Chemical Biological Center, Aberdeen Proving Ground, MD, 21010, USA.
Alvin T LiemU.S. Army Combat Capabilities Development Command Chemical Biological Center, Aberdeen Proving Ground, MD, 21010, USA.
Henry S GibbonsU.S. Army Combat Capabilities Development Command Chemical Biological Center, Aberdeen Proving Ground, MD, 21010, USA. henry.s.gibbons.civ@mail.mil.ORCID http://orcid.org/0000-0002-5603-8679

Funding

Defense Threat Reduction Agency CB3654
6 · The paper itself

Abstract

backgroundTracking dispersal of microbial populations in the environment requires specific detection methods that discriminate between the target strain and all potential natural and artificial interferents, including previously utilized tester strains. Recent work has shown that genomic insertion of short identification tags, called "barcodes" here, allows detection of chromosomally tagged strains by real-time PCR. Manual design of these barcodes is feasible for small sets, but expansion of the technique to larger pools of distinct and well-functioning assays would be significantly aided by software-guided design.

resultsHere we introduce barCoder, a bioinformatics tool that facilitates the process of creating sets of uniquely identifiable barcoded strains. barCoder utilizes the genomic sequence of the target strain and a set of user-specified PCR parameters to generate a list of suggested barcode "modules" that consist of binding sites for primers and probes, and appropriate spacer sequences. Each module is designed to yield optimal PCR amplification and unique identification. Optimal amplification includes metrics such as ideal melting temperature and G+C content, appropriate spacing, and minimal stem-loop formation; unique identification includes low BLAST hits against the target organism, previously generated barcode modules, and databases (such as NCBI). We tested the ability of our algorithm to suggest appropriate barcodes by generating 12 modules for Bacillus thuringiensis serovar kurstaki-a simulant for the potential biowarfare agent Bacillus anthracis-and three each for other potential target organisms with variable G+C content. Real-time PCR detection assays directed at barcodes were specific and yielded minimal cross-reactivity with a panel of near-neighbor and potential contaminant materials.

conclusionsThe barCoder algorithm facilitates the generation of synthetically barcoded biological simulants by (a) eliminating the task of creating modules by hand, (b) minimizing optimization of PCR assays, and (c) reducing effort wasted on non-unique barcode modules.

Indexed as

Bacillus anthracisDNA Barcoding, TaxonomicDNA PrimersAlgorithmsGenomeReal-Time Polymerase Chain ReactionDNA PrimersDNA barcodesGenetic barcodingGenome taggingMicrobial forensicsqPCR detectionTagged strains

Identifiers

PMID33648451
PMCPMC7919090

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.