Research Article
Full Length Research Paper
Comparison of three DNA extraction methods for polymerase chain reaction (PCR) analysis of bacterial genomic DNA
*Ibrahim Al-Hsawi, Abdul Al-Habib and Mohammed Hakami
Department of Environmental and Health Research, Faculty of Medicine, King Faisal University, Hofuf, Saudi Arabia.
E-mail: [email protected]
Accepted 26 September, 2014
Abstract
Rapid isolation of high-purity microbial genomic DNA is necessary for genome analysis. In this study, the authors compared a one-hour procedure using a microwave with enzymatic and boiling methods of genomic DNA extraction from Gram-negative and Gram-positive bacteria. High DNA concentration and purity were observed for both MRSA and ESBL strains (80.1 and 91.1 µg/ml; OD260/280, 1.82 and 1.70, respectively) when the extraction protocol included microwave pre-heating. DNA quality was further confirmed by PCR detection of mecA and CTX-M. In conclusion, the microwave-based procedure was rapid, efficient, cost-effective, and applicable for both Gram-positive and Gram-negative bacteria.
Key words: DNA, MRSA, ESBL, gene, enzymatic, boiling, microwave.
*Ibrahim Al-Hsawi, Abdul Al-Habib and Mohammed Hakami
Page: 23 - 26
Research Article
African Journal of DNA and Cell Biology Vol. 1 (1) pp. 009-022, October, 2014. © International Scholars Journals
Full Length Research Paper
Location of anti-infection resistance genes of Escherichia coli from residential livestocks in south east Nigeria with DNA microarray
Nnaji W. Ngozi
Department of Biotechnology, Faculty of Medical Sciences, University of Port-Harcourt, Rivers, Nigeria.
Email:[email protected]
Accepted 18 September, 2014
Abstract
DNA microarray was developed for detection of up to 90 antibiotic resistance genes in Escherichia coli by hybridization. Each antibiotic resistance gene was represented by two specific oligonucleotides chosen from consensus sequences of gene families. A total of 203 oligonucleotides (50-100 base) were spotted onto the microarray. The sequence identity of each gene was compared with GenBank sequen-ces, biotin was used as the positive control and 16s rRNA as orientation. Of the 40 E. coli isolates analyzed in this study, 37 were identified as having, at least, one antibiotic resistance gene. Among the different antibiotic resistance genes detected, bla-CMY-2 and strA were the most prevalent occurring in 28 (70%) of the isolates, respectively. Other common genes included were TEM1 11(27.5%), Sul2 14 (35%) and TetA 21(52.5%). The microarray genotyping corresponded with the phenotype of the strains. The disposable microarray presents the advantage of rapidly screening bacteria for the pre-sence of known antibiotic resistance genes. This technology has a large potential for applications in basic research, food safety, and surveillance programs for antimicrobial resistance.
Key words: DNA microarray, antibiotic resistance, Escherichia coli.
Nnaji W. Ngozi
Page: 9 - 22
Research Article
African Journal of DNA and Cell Biology Vol. 1 (1) pp. 001-008, October, 2014. © International Scholars Journals
Full Length Research Paper
Hereditary differences in Nigerian brinjal eggplant (Solanum melongena L.) as uncovered by arbitrary intensified polymorphic DNA (RAPD) markers
B. J Ayodeji1,3*, Iroruoture Cynthia1, Oguname Oscar3, Omonode J. E1, Olarutekpu O. Tony1, , Ihemefuna Bolaji1 and Temitope E. George2
1Department of Cell Biology and Genetics, Faculty of Biological Sciences, University of Ibadan, Ibadan, Nigeria.
2Department of Botany, Faculty of Medicine, Imo State University, Owerri, Nigeria.
3Department of Molecular Biology, Faculty of Biotechnology, Rivers State University of Science and Technology, Port Harcourt, Nigeria.
E-mail: [email protected]
Accepted 8 September, 2014
Abstract
The taxonomy of Solanum melongena L., also known as brinjal eggplant, has remained difficult because previous studies to establish genetic relationships among taxa are mainly based on morphological features, which are insufficient to establish genetic affinities. In the present investigation, five highly polymorphic random amplified polymorphic DNA primers were used to describe the genetic similarity and diversity among its accessions in Nigeria. The results show a high level of polymorphism based on the banding patterns among the samples. This indicated a wide and diverse genetic base. Four distinct clusters were equally noticeable at a coefficient of 0.80 from the dendrogram generated. Generally, the grouping pattern clearly indicates that irrespective of variations in fruit traits among samples, they were still grouped together in different clusters with a high similarity coefficient value. This probably showed some genetic relatedness/closeness among the samples concerned. The study also revealed that there is no association between RAPD pattern and the geographic origin of accessions. These agreed with previously published data on the characterization of eggplant. The study disclosed that molecular methods coupled with morphological analyses could make proper classification of S. melongena and other Solanum species in Nigeria possible to achieve.
Key words: Eggplant, random amplified polymorphic DNA (RAPD), Data, polymorphism, taxonomy.
Iroruoture Cynthia, B. J Ayodeji*, Olarutekpu O. Tony, Omonode J. E, Ihemefuna Bolaji and Temitope E. George, Oguname Oscar
Page: 1 - 8