DNA barcodes with short sequence fragments are used for species identification. Because of advances in sequencing technologies, DNA barcodes have gradually been emphasized. DNA sequences from different organisms are easily and rapidly acquired. Therefore, DNA sequence analysis tools play an increasingly crucial role in species identification. This study proposed deep barcoding, a deep learning framework for species classification by using DNA barcodes. Deep barcoding uses raw sequence data as the input to represent one-hot encoding as a one-dimensional image and uses a deep convolutional neural network with a fully connected deep neural network for sequence analysis. It can achieve an average accuracy of >90 percent for both simulation and real datasets. Although deep learning yields outstanding performance for species classification with DNA sequences, its application remains a challenge. The deep barcoding model can be a potential tool for species classification and can elucidate DNA barcode-based species identification.

DNA barcoding is widely used in fields, such as taxonomy and species identification. Conventional DNA barcoding sequences employ uninformative or repeat nucleotides in known groups of taxa within a monophylum. Herein, we propose a decision theory-based DNA barcode that tests for the ribulose bisphosphate carboxylase gene (rbcL). The proposed method can generate shorter DNA barcodes called single nucleotide polymorphism (SNP) tags, which shorten rbcL sequences from their full length (400-654 bp) to 25-bp DNA tags. These DNA tags are then represented by quick response (QR) codes containing the species names, accession numbers, and DNA tag sequences. Our proposed method can efficiently reduce data storage and provide DNA barcoding for various plant species.

Many drug or single nucleotide polymorphism (SNP)-related resources and tools have been developed, but connecting and integrating them is still a challenge. Here, we describe a user-friendly web-based software package, named Drug-SNPing, which provides a platform for the integration of drug information (DrugBank and PharmGKB), protein-protein interactions (STRING), tagSNP selection (HapMap) and genotyping information (dbSNP, REBASE and SNP500Cancer). DrugBank-based inputs include the following: (i) common name of the drug, (ii) synonym or drug brand name, (iii) gene name (HUGO) and (iv) keywords. PharmGKB-based inputs include the following: (i) gene name (HUGO), (ii) drug name and (iii) disease-related keywords. The output provides drug-related information, metabolizing enzymes and drug targets, as well as protein-protein interaction data. Importantly, tagSNPs of the selected genes are retrieved for genotyping analyses. All drug-based and protein-protein interaction-based SNP genotyping information are provided with PCR-RFLP (PCR-restriction enzyme length polymorphism) and TaqMan probes. Thus, users can enter any drug keywords/brand names to obtain immediate information that is highly relevant to genotyping for pharmacogenomics research.

Genetic association is a challenging task for the identification and characterization of genes that increase the susceptibility to common complex multifactorial diseases. To fully execute genetic studies of complex diseases, modern geneticists face the challenge of detecting interactions between loci. A genetic algorithm (GA) is developed to detect the association of genotype frequencies of cancer cases and noncancer cases based on statistical analysis. An improved genetic algorithm (IGA) is proposed to improve the reliability of the GA method for high-dimensional SNP-SNP interactions. The strategy offers the top five results to the random population process, in which they guide the GA toward a significant search course. The IGA increases the likelihood of quickly detecting the maximum ratio difference between cancer cases and noncancer cases. The study systematically evaluates the joint effect of 23 SNP combinations of six steroid hormone metabolisms, and signaling-related genes involved in breast carcinogenesis pathways were systematically evaluated, with IGA successfully detecting significant ratio differences between breast cancer cases and noncancer cases. The possible breast cancer risks were subsequently analyzed by odds-ratio (OR) and risk-ratio analysis. The estimated OR of the best SNP barcode is significantly higher than 1 (between 1.15 and 7.01) for specific combinations of two to 13 SNPs. Analysis results support that the IGA provides higher ratio difference values than the GA between breast cancer cases and noncancer cases over 3-SNP to 13-SNP interactions. A more specific SNP-SNP interaction profile for the risk of breast cancer is also provided.

The polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique is often used in laboratories and many basic research studies of complex genetic diseases associated with single nucleotide polymorphisms (SNP). When performing PCR-RFLP for SNP genotyping, feasible primer pairs are subject to numerous constraints and require a restriction enzyme for discriminating the target SNP. In this study, we develop a method for natural PCR-RFLP primer design for SNP genotyping using a particle swarm optimization algorithm. The in silico simulation with SNPs of the SLC6A4 gene demonstrates that this method reliably produces designs for natural PCR-RFLP primers which best fit the common primer constraints and also identifies available restriction enzymes.

Many association studies analyze the genotype frequencies of case and control data to predict susceptibility to diseases and cancers. An increasing number of studies has shown that the risk of getting diseases and cancers is associated with the co-occurrence of some contain single nucleotide polymorphisms (SNPs). Determining the disease-causing SNPs has become an important objective. In order to study the SNP-SNP interaction in breast cancer, we used a particle swarm optimization (PSO) algorithm to compute the difference between the control and case data and performed a feature selection from different SNP combinations with their corresponding genotypes. The best combination of SNP-SNP interactions is the maximal difference of co-occurrences between the control and case groups. In this study, we explored the SNP interaction of 19 SNPs in 372 controls and 398 cases of breast cancer association using simulated SNP data of breast cancers. The odds ratio (OR) were used to evaluate the breast cancer risk in terms of the best combination of SNP-SNP interactions. Compared to their corresponding non-SNP combinations, the estimated OR of the best predicted SNP combination with genotypes for breast cancer is significantly greater than 1 (about 1.771 and 2.417; confidence interval (CI): 1.223-4.371; p <; 0.05-0.001) for specific SNP combinations of two to five SNPs. The SNP interaction associated with a high risk of breast cancer could be successfully predicted using the proposed PSO method.

BACKGROUND: PCR-restriction fragment length polymorphism (RFLP) assay is a cost-effective method for SNP genotyping and mutation detection, but the manual mining for restriction enzyme sites is challenging and cumbersome. Three years after we constructed SNP-RFLPing, a freely accessible database and analysis tool for restriction enzyme mining of SNPs, significant improvements over the 2006 version have been made and incorporated into the latest version, SNP-RFLPing 2. RESULTS: The primary aim of SNP-RFLPing 2 is to provide comprehensive PCR-RFLP information with multiple functionality about SNPs, such as SNP retrieval to multiple species, different polymorphism types (bi-allelic, tri-allelic, tetra-allelic or indels), gene-centric searching, HapMap tagSNPs, gene ontology-based searching, miRNAs, and SNP500Cancer. The RFLP restriction enzymes and the corresponding PCR primers for the natural and mutagenic types of each SNP are simultaneously analyzed. All the RFLP restriction enzyme prices are also provided to aid selection. Furthermore, the previously encountered updating problems for most SNP related databases are resolved by an on-line retrieval system. CONCLUSIONS: The user interfaces for functional SNP analyses have been substantially improved and integrated. SNP-RFLPing 2 offers a new and user-friendly interface for RFLP genotyping that can be used in association studies and is freely available at http://bio.kuas.edu.tw/snp-rflping2.

BACKGROUND: Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) method produces allele-specific DNA bands of different lengths by adding four designed primers and it achieves the single nucleotide polymorphism (SNP) genotyping by electrophoresis without further steps. It is a time- and cost-effective SNP genotyping method that has the advantage of simplicity. However, computation of feasible CTPP primers is still challenging. RESULTS: In this study, we propose a GA (genetic algorithm)-based method to design a feasible CTPP primer set to perform a reliable PCR experiment. The SLC6A4 gene was tested with 288 SNPs for dry dock experiments which indicated that the proposed algorithm provides CTPP primers satisfied most primer constraints. One SNP rs12449783 in the SLC6A4 gene was taken as an example for the genotyping experiments using electrophoresis which validated the GA-based design method as providing reliable CTPP primer sets for SNP genotyping. CONCLUSIONS: The GA-based CTPP primer design method provides all forms of estimation for the common primer constraints of PCR-CTPP. The GA-CTPP program is implemented in JAVA and a user-friendly input interface is freely available at http://bio.kuas.edu.tw/ga-ctpp/.

Many single nucleotide polymorphisms (SNPs) genotyping techniques have been developed but most of them are expensive. Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) is a restriction enzyme-free and economic genotyping but its primer design is still computationally challenged. Here, we introduced a genetic algorithm (GA)-based PCR-CTPP primer design method. Thirty SNPs of the Janus kinase 2 gene with their SNP flanking length for 500 bps were tested. These GA-based designing CTPP primers were characterized with close values for melting temperature (Tm) and specificity, and their corresponding PCR products were provided with the optimal length. In conclusion, this novel PCR-CTPP primer designing method provides the computation for the primer information of cost- and time-effective enzyme-free SNP genotyping.

A fuzzy system with dynamically adapt the inertia weight of the particle swarm optimization (FAPSO) had been implemented to select a specific feasible primer pair for PCR experiments. Overall, fifty accession nucleotide sequences between 1900 bps and 2100 bps were sampled for primer design with specific PCR product lengths of 150~300 bps and 500~800 bps. Total five hundred runs of the proposed primer design approaches were performed for each accession nucleotide sequence to calculate the optimum accuracy. The proposed approach is compared to standard PSO primer design method. The results generated in a dry dock experiment showed that the FAPSO primer design yielded approximately optimal primer sets and had a relatively short CPU-time than standard PSO. Related materials are available online at http://bio.kuas.edu.tw/fapso-pd/.

BACKGROUND: Mitochondrial single nucleotide polymorphisms (mtSNPs) constitute important data when trying to shed some light on human diseases and cancers. Unfortunately, providing relevant mtSNP genotyping information in mtDNA databases in a neatly organized and transparent visual manner still remains a challenge. Amongst the many methods reported for SNP genotyping, determining the restriction fragment length polymorphisms (RFLPs) is still one of the most convenient and cost-saving methods. In this study, we prepared the visualization of the mtDNA genome in a way, which integrates the RFLP genotyping information with mitochondria related cancers and diseases in a user-friendly, intuitive and interactive manner. The inherent problem associated with mtDNA sequences in BLAST of the NCBI database was also solved. DESCRIPTION: V-MitoSNP provides complete mtSNP information for four different kinds of inputs: (1) color-coded visual input by selecting genes of interest on the genome graph, (2) keyword search by locus, disease and mtSNP rs# ID, (3) visualized input of nucleotide range by clicking the selected region of the mtDNA sequence, and (4) sequences mtBLAST. The V-MitoSNP output provides 500 bp (base pairs) flanking sequences for each SNP coupled with the RFLP enzyme and the corresponding natural or mismatched primer sets. The output format enables users to see the SNP genotype pattern of the RFLP by virtual electrophoresis of each mtSNP. The rate of successful design of enzymes and primers for RFLPs in all mtSNPs was 99.1%. The RFLP information was validated by actual agarose electrophoresis and showed successful results for all mtSNPs tested. The mtBLAST function in V-MitoSNP provides the gene information within the input sequence rather than providing the complete mitochondrial chromosome as in the NCBI BLAST database. All mtSNPs with rs number entries in NCBI are integrated in the corresponding SNP in V-MitoSNP. CONCLUSION: V-MitoSNP is a web-based software platform that provides a user-friendly and interactive interface for mtSNP information, especially with regard to RFLP genotyping. Visual input and output coupled with integrated mtSNP information from MITOMAP and NCBI make V-MitoSNP an ideal and complete visualization interface for human mtSNPs association studies.