DNA sequencing is one of the most important techniques in the field of molecular biology, according to which the order of nucleotides can be determined in a piece of DNA. There are several different methods for DNA sequencing. Today, with advances in the field Nanotechnology and bioinformatics, methods have developed to increase the speed and efficiency of DNA sequencing, which has made it possible to carry out large projects such as sequencing the human genome cheaper, more accurately and more quickly. Today, DNA sequencing is an integral part of modern science. This technique is used in various fields of microbiology, including tracking infectious diseases and studying the diversity of microbial communities. There are different ways to sequence DNA. But each of these techniques has its advantages and disadvantages. To get acquainted with these methods, This article is a review that analyzes the generations of sequencing from the past to the present that has been collected from articles, reputable book sites and articles.

Hereditary hemolytic anemias present a unique diagnostic challenge due to their wide phenotypic and genotypic spectrum. Accurate diagnosis is essential to ensure appropriate treatment. We report two cases, which presented as hemolytic anemias, but initial workup was inconclusive and they were finally diagnosed with the help of Next Generation Sequencing (Dehydrated Hereditary Stomatocytosis and Kӧ ln Hemoglobinopathy). The introduction of gene sequencing to aid diagnosis of these disorders is a revolutionary step forward and should be incorporated earlier in the workup of such patients...

miRNA genes consider as an important class of gene expression regulators that have diverse responses to environmental stresses and play a key role in regulating gene expression during post-transcriptional regulation. Hypericum perforatum, is well-known medicinal plant for its application in the treatment of depression due to the effects of its bioactive compounds named hypericin and hyperforin. The present study was performed to identify the conserved miRNAs and their target genes in the transcriptome of H. perforatum. First, RNA-seq data were obtained from the European Nucleotide Archive (ENA) of EMBL-EBI database and then the transcriptome (RNA) sequences were assembled. The non-coding transcripts were identified and considered as miRNA precursors candidate sequences. Finally, six miRNAs named Hp-miR395, Hp-miR845d, Hp-miR414, Hp-miR159, Hp-miR159e, and Hp-miR156c were identified from the candidate sequences using of C-mii software after applying strict filters.  Investigation of the protein-protein interaction network determined the relationships between the target genes and target proteins especially in transcription factors. In the next step, to confirm the target genes for the identified miRNAs, the foliar application of Methyl Jasmonate (MJ) with concentrations of 0 (control) and 200 μM was performed on H. perforatum plants and the expression pattern of two target genes was investigated. In the analysis of qRT-PCR expression changes at 12, 24, 48 and 72 hours after applying MJ, the relative expression level of Hyp-1 (DN121523_c1_g4_i2) and HD-Zip (DN121003_c0_g1_i1) transcripts increased after 72 hours of application of MJ. In general, regarding to the regulatory role of the identified miRNAs in the present study, these genes could be used to better and more accurately identify the biosynthetic pathway of hypercin and hyperforin.

Beta-thalassemia is a group of inherited blood disorders with autosomal recessive inheritance. The most effective strategy is to reduce the incidence of this genetic disease, which can be accomplished by utilizing appropriate prenatal diagnosis in clinical practice. Aggressive biopsy during the pregnancy is associated with an abortion risk of approximately 1% for the fetus. Free fetal DNA in maternal plasma is an excellent source of genetic material for prenatal molecular diagnoses. Relative haplotype dosage (RHDO), a haplotype‐ based approach, has shown promise as an application for noninvasive prenatal diagnosis. This study was conducted to investigate beta thalassemiain the fetus through maternal blood with NGS technique and RHDO analysis. Total of 5 beta-thalassemia carrier couples (minor) were genotyped by ARMS-PCR for IVSII-I G>A mutation. During the pregnancy, 10 ml of blood was collected from pregnant women, A sample of saliva was also taken from their Husbands. Samples were sent to the Genoma Center of Italy for NGS examination. Finally, results were compared with those of the invasion method. The results of NGS technique with RHDO analysis without having to analyze the affected person by haplotypic block analysis have 100% accuracy. The results with cffDNA were confirmed by invasive methods according to the national protocol for beta-thalassemia. A total of 5 fetal samples belong to above couples were genotyped for IVSII-I G>A mutation, in which 1 fetus were affected, 2 fetus were normal and 2 fetus were carrier of beta-thalassemia. Sensitivity and specificity of NGS test were equal to 100% and 100% respectively. Also, positive and negative predictive values were obtained as 100% and 100%, respectively. Targeted sequencing of maternal plasma DNA for NIPD of beta-thalassemia and using SHAPEIT software for RHDO analysis is feasible, proved that it is applicable to construct parental haplotypes without information from other family members.

Today, medicinal plants are used in the treatment of many diseases because of their secondary metabolites. Thyme as one of these plants contains a wide range of secondary metabolites such as terpenes. Various methods have been developed to increase these materials. In classical methods, environmental factors are changed to produce the most effective substance in medicinal plants, but in newer approaches that are based on plant genetics, higher yields are observed. One of these approaches is the use of miRNAs. These miRNAs control gene expression after transcription by mRNA analysis or inhibition of their translation, and play different roles in biological and metabolic processes in plants and animals. One of the simplest and least expensive methods for identifying miRNAs is the use of bioinformatics tools and methods. To identify distinct miRNA in different species of thyme, a study based on homology search was conducted using transcriptomic data of thyme. First, this information was refined and then aligament performed against all known miRNAs in the miRBase database. After screening of results based on factors such as length and e-value level, the secondary structure of miRNAs was analyzed with UNAfold tool. Target genes were identified using psRNATARGET tool and phylogenetic relationships were investigated using maximum likelihood method and RaxML tool. In total 64 distinct candidate’s miRNAs were identified in different species of thymus and 14 miRNAs included miR172 and miR396 played an important role in terpenes synthesis and it has been proven in previous studies. The phylogenetic tree was able to show the relationship between miRNAs in different species.

Background: A lizard Leishmania has been isolated from a lizard (Agama agilis) in Iran. Its genome sequence has not been determined, so far. Methods: The study was done at Shahid Beheshti University of Medical Sciences, Tehran, Iran in 2017-2023. Leishmania promastigotes were cultured in RPMI1640 culture medium and collected at logarithmic phase by centrigugation. Parasite RNA was extracted by the Qiagene standard kit and its quantity and quality was determined and sequenced by NGS method with Illumina PE machine at BGI Company (China). Results: The number of 8316 mRNA, 83 tRNA, 63 rRNA, 83 ncRNA, 5 snRNA, 1039 snoRNA, 36 region, and 3 repeat regions, 8343 CDS, 9597 Exon and 9292 Genes were identified in promastigote of Iranian lizard Leishmania. Conclusion: Genomic elements of Iranian lizards Leishmania (with unique characteristics) were determined and identified by NGS system.

Cancer is one of the leading causes of death worldwide and one of the greatest challenges in extending life expectancy. The paradigm of one-size-fits-all medicine has already given way to the stratification of patients by disease subtypes, clinical characteristics, and biomarkers (stratified medicine). The introduction of next-generation sequencing (NGS) in clinical oncology has made it possible to tailor cancer patient therapy to their molecular profiles. NGS is expected to lead the transition to precision medicine (PM), where the right therapeutic approach is chosen for each patient based on their characteristics and mutations. Here, we highlight how the NGS technology facilitates cancer treatment. In this regard, first, precision medicine and NGS technology are reviewed, and then, the NGS revolution in precision medicine is described. In the sequel, the role of NGS in oncology and the existing limitations are discussed. The available databases and bioinformatics tools and online servers used in NGS data analysis are also reviewed. The review ends with concluding remarks.