scholarly journals PUNS: transcriptomic- and genomic-in silico PCR for enhanced primer design

2004 ◽  
Vol 20 (15) ◽  
pp. 2399-2400 ◽  
Author(s):  
P. C. Boutros ◽  
A. B. Okey
Keyword(s):  
In Silico ◽  
Primer Design ◽  
In Silico Pcr

scholarly journals DETECTION OF VIRULENCE GENES phoP AND phoQ IN Salmonella spp. USING IN SILICO POLYMERASE CHAIN REACTION

2020 ◽  
Vol 4 (1) ◽  
Author(s):  
Stalis Norma Ethica ◽  
Hayatun Fuad ◽  
Nur Hidayah ◽  
Sri Sinto Dewi ◽  
Aditya Rahman Ernanto ◽  
...  
Keyword(s):  
In Silico ◽  
Salmonella Enterica ◽  
Virulence Genes ◽  
Gene Detection ◽  
Chain Reaction ◽  
Salmonella Spp ◽  
Detection Tool ◽  
In Silico Study ◽  
Polymerase Chain ◽  
In Silico Pcr

Detection of Salmonella bacteria based on their virulence genes is among essential steps in the eradication of clinical infection by bacteria. In this study, two pair of primers, PhoPF-PhoPR: 5’- CCGCGCAGGAAAAACTCAAA-3’ and 5’-ATCTGTTCCAGCATCACCGG -3’ as well as PhoQF-PhoQR: 5’-AGAGATGATGCGCGTACTGG-3’ and 5’- CAGACGCCCCATGAGAACAT-3’, had been successfully designed using Primer3Plus to detect the presence of phoP and phoQ genes in Salmonella spp. Using genomic DNA of 44 genomic data of Salmonella spp. as templates, PhoPF-PhoPR could produce 520-bp amplicon, while PhoQF-PhoQR could result in 598-bp amplicon. Results of in silico PCR showed that both pairs of primers PhoPF-PhoPR and PhoQF-PhoQR could detect only Salmonella enterica species, and no Salmonella bongori species could be detected based on phoP and phoQ sequences. Both pairs of PhoPF-PhoPR and PhoQF-PhoQR primers were also able to detect the virulence genes in most of the studied subspecies of Salmonella enterica available in silico database unless Arizona subspecies. As conclusion, based on this in silico study, phoP and phoQ genes appeared to be biomarkers for Salmonella enterica species. Both pairs of primers designed in this study has potential to be used as detection tool to differentiate species Salmonella enterica from Salmonella bongori, and also to distinguish S.enterica subsp. enterica from subsp. Arizonae.Keywords: Gene detection, bacterial virulence, phoP, phoQ, Salmonella spp.

Primer Design, Evaluation of Primer Universality, and Estimation of Identification Power of Amplicon Sequences In Silico

2019 ◽  
pp. 21-36
Author(s):  
Akifumi S. Tanabe ◽  
Satoshi Nagai ◽  
Yuki Hongo ◽  
Motoshige Yasuike ◽  
Yoji Nakamura ◽  
...  
Keyword(s):  
In Silico ◽  
Primer Design ◽  
Design Evaluation

scholarly journals Distribution of Biofilm-associated Genes among Acinetobacter baumannii by in-silico PCR

2021 ◽  
pp. 140-149
Author(s):  
A. Aldrin Joshua ◽  
A. S. Smiline Girija ◽  
P. Sankar Ganesh ◽  
J. Vijayashree Priyadharsini
Keyword(s):  
Acinetobacter Baumannii ◽  
In Silico ◽  
Target Genes ◽  
Multidrug Resistant ◽  
Pcr Analysis ◽  
Evolutionary Analysis ◽  
Ompa Gene ◽  
Clinical Strains ◽  
Tract Infections ◽  
In Silico Pcr

Background: Acinetobacter baumannii is a coccobacillus that is Gram negative, non motile, non fermentative and oxidase negative. It is the most common and successful nosocomial pathogen recognised by WHO. This dreadful pathogen causes urinary tract infections, ventilator associated pneumonia (VAP), bacteremia, etc., These infections are most common in hospital wards especially Intensive Care Unit (ICU). The infections are due to biofilm formation by the virulent genes of A. baumannii, and the common biofilm-associated genes of A. baumannii were bap, csuE, fimH, epsA, bfmS, ptk, pgaB, ompA, blaPER-1. Among these, bap, epsA and ompA genes are highly prevalent among the clinical strains of A. baumannii. Aim:  To detect the three vital biofilm-associated genes of A. baumannii by in-silico PCR analysis. Materials and Methods: 19 isolates of A. baumannii were selected and 3 target genes, namely epsA, ompA and bap gene were used for the amplification process through in-silico PCR simulation tools. Evolutionary analysis was done for the ompA gene. Results: The epsA gene was expressed in 10.52% of the total strains selected with the highest occurrence of ompA gene as 57.89%. bap gene was not observed from the study strains included. From evolutionary analysis based on ompA distributed strains, the Acinetobacter baumannii SDF and Acinetobacter baumannii BJAB0715 might be the parental strains where the evolution of strains would have started. Through successive generations, the Acinetobacter baumannii MDR-ZJ06 and Acinetobacter baumannii TYTH-1 had become the multidrug resistant strains present in the environment. Conclusion: The findings of the study confirms the distribution of epsA and ompA genes among the 19 different strains of A. baumannii. The study suggests periodical monitoring of biofilm based virulence genes among the clinical strains and to curtail the A. baumannii infections.

scholarly journals KASPspoon: an in vitro and in silico PCR analysis tool for high-throughput SNP genotyping

2019 ◽  
Vol 35 (17) ◽  
pp. 3187-3190 ◽  
Author(s):  
Alsamman M Alsamman ◽  
Shafik D Ibrahim ◽  
Aladdin Hamwieh
Keyword(s):  
High Throughput ◽  
In Silico ◽  
Genetic Maps ◽  
Supplementary Information ◽  
Pcr Analysis ◽  
Analysis Tool ◽  
Data Types ◽  
Kasp Assay ◽  
In Silico Pcr

Abstract Motivation Fine mapping becomes a routine trial following quantitative trait loci (QTL) mapping studies to shrink the size of genomic segments underlying causal variants. The availability of whole genome sequences can facilitate the development of high marker density and predict gene content in genomic segments of interest. Correlations between genetic and physical positions of these loci require handling of different experimental genetic data types, and ultimately converting them into positioning markers using a routine and efficient tool. Results To convert classical QTL markers into KASP assay primers, KASPspoon simulates a PCR by running an approximate-match searching analysis on user-entered primer pairs against the provided sequences, and then comparing in vitro and in silico PCR results. KASPspoon reports amplimers close to or adjoining genes/SNPs/simple sequence repeats and those that are shared between in vitro and in silico PCR results to select the most appropriate amplimers for gene discovery. KASPspoon compares physical and genetic maps, and reports the primer set genome coverage for PCR-walking. KASPspoon could be used to design KASP assay primers to convert QTL acquired by classical molecular markers into high-throughput genotyping assays and to provide major SNP resource for the dissection of genotypic and phenotypic variation. In addition to human-readable output files, KASPspoon creates Circos configurations that illustrate different in silico and in vitro results. Availability and implementation Code available under GNU GPL at (http://www.ageri.sci.eg/index.php/facilities-services/ageri-softwares/kaspspoon). Supplementary information Supplementary data are available at Bioinformatics online.

scholarly journals Assessment of sequence variation from SIT1 gene on diatom (Bacillariophyceae) using in silico PCR

2016 ◽  
Author(s):  
Immanuel Sanka ◽  
Niken Satuti Nur Handayani ◽  
Eko Agus Suyono ◽  
Riza Arief Putranto
Keyword(s):  
In Silico ◽  
Sequence Variation ◽  
In Silico Pcr

scholarly journals PrimerMiner: an R package for development and in silico validation of DNA metabarcoding primers

2016 ◽  
Author(s):  
Vasco Elbrecht ◽  
Florian Leese
Keyword(s):  
In Silico ◽  
Sequence Data ◽  
Dna Barcode ◽  
R Package ◽  
Primer Design ◽  
Reference Sequence ◽  
Gene Marker ◽  
Sequence Alignments ◽  
Variable Binding ◽  
Dna Metabarcoding

1) DNA metabarcoding is a powerful tool to assess biodiversity by amplifying and sequencing a standardized gene marker region. Its success is often limited due to variable binding sites that introduce amplification biases. Thus the development of optimized primers for communities or taxa under study in a certain geographic region and/or ecosystems is of critical importance. However, no tool for obtaining and processing of reference sequence data in bulk that serve as a backbone for primer design is currently available. 2) We developed the R package PrimerMiner, which batch downloads DNA barcode gene sequences from BOLD and NCBI databases for specified target taxonomic groups and then applies sequence clustering into operational taxonomic units (OTUs) to reduce biases introduced by the different number of available sequences per species. Additionally, PrimerMiner offers functionalities to evaluate primers in silico, which are in our opinion more realistic then the strategy employed in another available software for that purpose, ecoPCR. 3) We used PrimerMiner to download cytochrome c oxidase subunit I (COI) sequences for 15 important freshwater invertebrate groups, relevant for ecosystem assessment. By processing COI markers from both databases, we were able to increase the amount of reference data 249-fold on average, compared to using complete mitochondrial genomes alone. Furthermore, we visualized the generated OTU sequence alignments and describe how to evaluate primers in silico using PrimerMiner. 4) With PrimerMiner we provide a useful tool to obtain relevant sequence data for targeted primer development and evaluation. The OTU based reference alignments generated with PrimerMiner can be used for manual primer design, or processed with bioinformatic tools for primer development.

In Silico PCR Primer Designing and Validation

2015 ◽  
pp. 143-151 ◽  
Author(s):  
Anil Kumar ◽  
Nikita Chordia
Keyword(s):  
In Silico ◽  
In Silico Pcr ◽  
Pcr Primer
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