Background and Objectives The use of positive controls in Real-Time PCR experiments is crucial for ensuring assay accuracy and reliability. Traditionally, such controls are derived from clinical resources. However, in situations where access to biological samples is limited, Polymerase Cycle Assembly (PCA) provides a reliable alternative for creating positive controls. The aim of this study was to evaluate the efficiency of the enzymatic method for producing genomic DNA fragments suitable for molecular biology applications. To the best of our knowledge, while PCA has been primarily employed for assembling sequences, its utility in constructing HSV-2 positive controls has not yet been investigated. Materials and Methods In this experimental study, the first step involved designing overlapping DNA fragments targeting the US6 region of the HSV-2 genome, a unique and well-characterized sequence selected as a suitable target. The selection of the US6 region was based on a literature review analysis. The overlapping fragments were evaluated for secondary structures and thermodynamic parameters using AlleleID software. Primer design for the US6 region was performed using MEGA version 11, ensuring specificity to the HSV-2 genome. The evaluation of the genome HSV-2 with the SnapGene software has been carried out. A comprehensive examination of the primers and their products from the secondary structure was conducted using MFold. This software analyses the sequence intended for primer design in terms of its secondary structure. Results Alignment analysis revealed a high degree of sequence similarity between HSV-1 and HSV-2. Therefore, primers and overlapping fragments were designed to target a region exhibiting the greatest natural variation. The results demonstrated that the in silico positive controls were comparable to natural controls, and the specificity of designed primers was confirmed.   Using SnapGene software, the size of reproduced DNA segment was estimated to be approximately 70 base pairs, providing a suitable template for future studies. Conclusions  The results demonstrate that PCA is an effective and highly reliable method for synthesizing positive controls for molecular diagnostics. Targeting the HSV-2 US6 region ensured specificity and structural integrity of the synthetic fragment.

The purpose of this research is to deal with the problem of two-stage Assembly flow shop scheduling. A number of single-item products (identical) each formed of several different parts are ordered. Each part has m operations done at the first stage with m different machines. After manufacturing the parts, they are assembled into a final product with some non-identical machines. The purpose of the problem is to find the optimal sequence of the parts in the manufacturing stage, allocation and the optimal sequence of the products in the Assembly stage. A mixed integer linear programming model and two metaheuristic algorithms, which are particle swarm with local search (MPSO) and simulated annealing (SA), are presented to solve this problem. Computational experiments are conducted to evaluate the performance of the proposed model and algorithms. The results show that the MPSO algorithm performs better than the SA one.

Automobile sector forms the backbone of manufacturing sector. Vehicle Assembly line is important section in automobile plant where repetitive tasks are performed one after another at different workstations. In this thesis, a methodology is proposed to reduce cycle time and time loss due to important factors like equipment failure, shortage of inventory, absenteeism, set-up, material handling, rejection and fatigue to improve output within given cost constraints. Various relationships between these factors, corresponding cost and output are established by scientific approach. This methodology is validated in three different vehicle Assembly plants. Proposed methodology may help practitioners to optimize the Assembly line using lean techniques.

To assemble a variety of products, a multi single Assembly line, mixed model Assembly line or a flexible Assembly system (FAS) may be used. Past studies were concerned with the modeling, investigation and comparison of single and mixed model Assembly systems, where the main transportation system was a continuous conveyor. According to new market demands, it is required to consider FAS, utilizing AGVs as the main automatic transportation system. In this research, traditional and flexible Assembly systems are reviewed and compared. To compare these system criteria, such as stability to demand changes and balance of station workloads, the efficiency of material handling and the whole Assembly system are considered. The Assembly of two kinds of motorcycle is used as a sample case study. The result shows that a stable system (with little variation in terms of number of stations) having good performance measure values, is achieved when a flexible Assembly system is employed.