Mousazade Moghadam M, Babavalian H, Mirnejad R, Shakeri F,
Volume 6, Issue 1 (4-2012)
Abstract
Abstract
Background and objectives: Genomic DNA extraction of bacterial cells is of processes performed normally in most biological laboratories therefore, various methods have been offered, manually and kit, which may be time consuming and costly. In this paper, genomic DNA extraction of Staphylococcus aureus was investigated using some laundry detergent brands available in Iran to achieve a rapid and cost effective method.
Material and Methods: five-enzyme Taj brand, three-enzyme Saftlan brand ,and Darya and Pak brands without enzyme were used in the concentrations of 10, 20, 40, 80 mg/L. Afterwards, in order to evaluate the efficiency of extracted DNA in downstream processing, PCR test was performed for femA gene in the genome of Staphylococcus aureus.
Results: DNA extraction using different concentrations of the brands show that extracted DNA using 40 mg/L Saftlan and Taj brand powders have the best results according concentration (µg/ml) and purity (A260/A280) parameters. These parameters are 387.5 1.88 (Taj), 254.1 2.80 (Softlan), 396.6 1.95 (Manual) and 423.3 2.2 (Kit), respectively. Afterward, the PCR test results by show that DNA extraction using laundry detergents has no effect on its efficiency in order to be used in downstream processes.
Conclusion: These results indicate that the proper concentrations of laundry detergents can be used to extract genomic DNA with similar efficiency to kit and manual extraction methods.
Key words: Bacterial genome, DNA extraction, laundry powder, PCR, Staphylococcus aureus
Nima Shaykh Baygloo , Majid Bouzari , Fateh Rahimi ,
Volume 11, Issue 3 (5-2017)
Abstract
ABSTRACT
Background and Objective: Prophage sequences are major contributors to interstrain variations within the same bacterial species. Acinetobacter baumannii is a gram-negative bacterium that causes a wide range of nosocomial infections, especially in intensive care unit inpatients. Prophage sequences constitute a considerable proportion of several sequenced complete genomes of A. baumannii. The aim of this study was to investigate the presence of prophage sequences in A. baumannii strains isolated from burn patients, and compare the results with other studies.
Methods: Presence of eight prophage sequences was investigated in the genome of ten multi-drug resistant A. baumannii isolates obtained from burn sites of 10 burn patients in a hospital in Isfahan, Iran. PCR and sequencing were performed to detect the prophage sequences. The presence of the eight prophage sequences in the genome of A. baumannii strains from other studies was investigated by BLAST analysis of whole nucleotide sequence of prophage sequences.
Results: The isolates in the present study had different prophage sequence profiles. Two isolates did not contain any of the sequences, while two isolates contained three and two of the prophage sequences. Other isolates contained only one sequence. The prophage sequence profiles observed in this study were not found in A. baumannii isolates from other studies.
Conclusion: The results of this study indicate that the prophage sequences profile can be useful for studying the epidemiology of A. baumannii strains.
Keywords: Acinetobacter baumannii, genome, prophage sequences.
