scholarly journals Isolation and genomic amplification of fish and shrimps from mangrove ecosystems in North Sumatra

2021 ◽  
Vol 912 (1) ◽  
pp. 012033
Author(s):  
H Hairani ◽  
M Basyuni ◽  
A Susilowati
Keyword(s):  
Extraction Method ◽  
Pcr Amplification ◽  
Band Pattern ◽  
Genomic Amplification ◽  
Shrimp Species ◽  
16S Gene ◽  
Mangrove Ecosystems ◽  
Barcode Dna ◽  
High Level ◽  
North Sumatra

Abstract Mangrove is a very typical ecosystem with a high level of diversity of flora and fauna. However, mangrove ecosystems are also one of the most vulnerable locations to extinction due to the high disturbances that occur. Some of the species that are susceptible to such disturbances are fish and shrimp. The use of barcode DNA is considered a fairly effective way to evaluate the condition of fish and shrimp populations in North Sumatra. Therefore, this study aims to obtain information on the success of kit methods and amplification patterns of barcoding marks 16S for fish and shrimp in mangrove ecosystems North Sumatra. The results showed that the kit extraction method produced a pattern of band that varied thick, thin and even smear. PCR amplification results with the 16S gene showed a good band pattern and indicated that the 16S primer used can detect fish and shrimp species that are visualized in the form of DNA bands.

scholarly journals DNA extraction and pattern of crab and macrobentos from North Sumatran mangrove forest, Indonesia

2021 ◽  
Vol 912 (1) ◽  
pp. 012046
Author(s):  
A M Harahap ◽  
M Basyuni ◽  
I E Susetya
Keyword(s):  
Dna Extraction ◽  
Mangrove Forest ◽  
Morphological Analysis ◽  
Animal Species ◽  
Pcr Amplification ◽  
Sequencing Analysis ◽  
Chain Reaction ◽  
Mangrove Ecosystems ◽  
Polymerase Chain ◽  
North Sumatra

Abstract Mangrove forest ecosystem is one of the most productive and unique ecosystems that serves to protect coastal areas from various disturbances, as well as provide habitat for various animal species. The large number of crab species and macrobentos in mangrove ecosystems results in frequent errors in the naming of species that have similarities in morphological features. This problem can be solved through a comprehensive approach by combining morphological analysis with genetic analysi.This study aims to report a DNA extraction and PCR amplification prior was used for the identification of crab and macrozoobentos from mangrove forest, North Sumatra. Primer 16S has a conserved area so it is appropriately used in Polymerase Chain Reaction (PCR) and sequencing analysis to determine taxonomy, phylogeny and diversity between specie. Visualization of PCR amplification results with primer16S from crab samples and macrobentos resulting a low DNA band with a size of 206 bp and a high of 678bp

scholarly journals In Vitro Incorporation of Helicobacter pylori into Candida albicans Caused by Acidic pH Stress

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 489 ◽  
Author(s):  
Kimberly Sánchez-Alonzo ◽  
Cristian Parra-Sepúlveda ◽  
Samuel Vega ◽  
Humberto Bernasconi ◽  
Víctor L. Campos ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Candida Albicans ◽  
Pcr Amplification ◽  
Growth Curves ◽  
Acidic Ph ◽  
Ph Values ◽  
16S Gene ◽  
Wide Range ◽  
H Pylori

Yeasts can adapt to a wide range of pH fluctuations (2 to 10), while Helicobacter pylori, a facultative intracellular bacterium, can adapt to a range from pH 6 to 8. This work analyzed if H. pylori J99 can protect itself from acidic pH by entering into Candida albicans ATCC 90028. Growth curves were determined for H. pylori and C. albicans at pH 3, 4, and 7. Both microorganisms were co-incubated at the same pH values, and the presence of intra-yeast bacteria was evaluated. Intra-yeast bacteria-like bodies were detected using wet mounting, and intra-yeast binding of anti-H. pylori antibodies was detected using immunofluorescence. The presence of the H. pylori rDNA 16S gene in total DNA from yeasts was demonstrated after PCR amplification. H. pylori showed larger death percentages at pH 3 and 4 than at pH 7. On the contrary, the viability of the yeast was not affected by any of the pHs evaluated. H. pylori entered into C. albicans at all the pH values assayed but to a greater extent at unfavorable pH values (pH 3 or 4, p = 0.014 and p = 0.001, respectively). In conclusion, it is possible to suggest that H. pylori can shelter itself within C. albicans under unfavorable pH conditions.

scholarly journals The Influence of Accountability, Work Experience and Professionalism on The Quality of The Quality of The Work of Auditors

2020 ◽  
Vol 4 (3) ◽  
pp. 449-459
Author(s):  
Sihol Marito Simorangkir ◽  
Jonathan Dubery Bukit ◽  
Keumala Hayati
Keyword(s):  
Work Experience ◽  
Sampling Method ◽  
Financial Statements ◽  
The North ◽  
Sample Technique ◽  
Saturated Sample ◽  
High Level ◽  
North Sumatra ◽  
Regression Tests

The study was conducted aimed at examining the effect of accountability, work experience, and professionalism on the quality of auditor performance. auditors are required to have a high level of accountability because the auditor will present financial statements that must be accountable by himself. In addition to having a high accountability the auditor should also have good work experience so that the auditor can convey a logical and logical understanding of the errors contained in the financial statements. And auditors also require high professionalism so that auditors can carry out professional and good tasks. The object of research is the auditor who is working at the North Sumatra BPKP Office. The data is 70 respondents. The sampling method is the saturated sample technique. Analyze the data using validity, reliability and linear regression tests. And the results of the study prove that accountability has an influence on the quality of the work of the auditor, while work experience and professionalism do not have an influence on the quality of the work of the audiores.

scholarly journals Genetic Diversity of Some Sweet Cherry Cultivars Based on Molecular Markers

2016 ◽  
Vol 73 (2) ◽  
pp. 153
Author(s):  
Ioana Virginia Berindean ◽  
Elena Tămaş ◽  
Oana Maria Toderic ◽  
Ioan Zagrai
Keyword(s):  
Tree Species ◽  
Rapd Markers ◽  
Sweet Cherry ◽  
Prunus Avium ◽  
Molecular Level ◽  
Genetic Relationships ◽  
Pcr Amplification ◽  
High Level ◽  
Sweet Cherry Cultivars

Sweet cherry (Prunus avium L.), originated around the Caspian and Black Sea, is an important fruit tree species of economic interest, and hence, breeding and conservation are requested (. Genetic analysis at the molecular level can be used effectively to study molecular polymorphism existing between intraspecific and interspecific tree species and phylogenetic relationships between them and their hybrids. The purpose of this study was to characterize and determine genetic relationships among the sweet cherry native genotypes belonging to Fruit Research & Development Station Bistrita, Romania, using RAPD markers. To eliminate the existence of possible synonyms from national romanian collection, we collect four Van cultivars, from four different national collection. For molecular analysis of the 16 varieties of sweet cherry were considered 13 RAPD primers selected from the literature. They were later used to determine the genetic variability at the molecular level using PAST program, and the dendrogram was generated based on Jaccard’s genetic distance. The dendrogram constructed by PAST software. The quantity and quality of the DNA obtained was suitable to achieve PCR amplification step. Only seven out of the 13 RAPD primers have generate polymorphic bands. The rest of seven were monomorphics. The most polymorphic primer was OPB10 which generated 11 bands from which 100% were polymorphic.Seven RAPD primers generated a high level of polymorphism which allowed to divide these cherry varieties into two groups according to their genetic geographical origin and the pedigree.

scholarly journals A modified SDS-based DNA extraction method from raw soybean

2019 ◽  
Vol 39 (2) ◽  
Author(s):  
Yimiao Xia ◽  
Fusheng Chen ◽  
Yan Du ◽  
Chen Liu ◽  
Guanhao Bu ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Pcr Amplification ◽  
Isoamyl Alcohol ◽  
Monitoring Methods ◽  
Detection Techniques ◽  
Dna Yield ◽  
Seed Flour ◽  
Dna Extraction Method ◽  
Lysis Buffer

Abstract Soybean is the most important genetically modified (GM) oilseed worldwide. Regulations relating to the approval of biotech soybean varieties and product labeling demand accurate and reliable detection techniques to screen for GM soya. High-quality extracted DNA is essential for DNA-based monitoring methods. Thus, four widely used protocols (SDS, CTAB, DP305, and DNeasy Plant Mini Kit) were compared in the present study to explore the most efficient DNA extraction method for raw soya matrix. The SDS-based method showed the highest applicability. Then crucial factors influencing DNA yield and purity, such as SDS lysis buffer component concentrations and organic compounds used to isolate DNA, were further investigated to improve the DNA obtained from raw soybean seeds, which accounts for the innovation of this work. As a result, lysis buffer (2% SDS (w/v), 150 mM NaCl, 50 mM Tris/HCl, 50 mM EDTA, pH 8.0) and organic reagents including chloroform/isoamyl alcohol (24:1, v/v) (C: I), isopropanol, and ethanol corresponding to the extraction and first and second precipitation procedures, respectively, were used in the optimized SDS method. The optimized method was verified by extracting approximately 2020–2444 ng DNA/mg soybean with A260/280 ratios of 1.862–1.954 from five biotech and non-biotech soybean varieties. Only 0.5 mg of soya was required to obtain enough DNA for PCR amplification using the optimized SDS-based method. These results indicate that the screening protocol in the present study achieves the highest suitability and efficiency for DNA isolation from raw soya seed flour.

scholarly journals Genetic Characterization of Highly Fluoroquinolone-Resistant Clinical Escherichia coli Strains from China: Role ofacrR Mutations

2001 ◽  
Vol 45 (5) ◽  
pp. 1515-1521 ◽  
Author(s):  
Hui Wang ◽  
Joann L. Dzink-Fox ◽  
Minjun Chen ◽  
Stuart B. Levy
Keyword(s):  
Escherichia Coli ◽  
Blot Analysis ◽  
Genetic Basis ◽  
Efflux Pump ◽  
Pcr Amplification ◽  
Fluoroquinolone Resistance ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
E Coli ◽  
High Level

ABSTRACT The genetic basis for fluoroquinolone resistance was examined in 30 high-level fluoroquinolone-resistant Escherichia coliclinical isolates from Beijing, China. Each strain also demonstrated resistance to a variety of other antibiotics. PCR sequence analysis of the quinolone resistance-determining region of the topoisomerase genes (gyrA/B, parC) revealed three to five mutations known to be associated with fluoroquinolone resistance. Western blot analysis failed to demonstrate overexpression of MarA, and Northern blot analysis did not detect overexpression of soxS RNA in any of the clinical strains. The AcrA protein of the AcrAB multidrug efflux pump was overexpressed in 19 of 30 strains of E. colitested, and all 19 strains were tolerant to organic solvents. PCR amplification of the complete acrR (regulator/repressor) gene of eight isolates revealed amino acid changes in four isolates, a 9-bp deletion in another, and a 22-bp duplication in a sixth strain. Complementation with a plasmid-borne wild-type acrR gene reduced the level of AcrA in the mutants and partially restored antibiotic susceptibility 1.5- to 6-fold. This study shows that mutations in acrR are an additional genetic basis for fluoroquinolone resistance.

scholarly journals Novel Tetracycline Resistance Gene,tet(32), in the Clostridium-Related Human Colonic Anaerobe K10 and Its Transmission In Vitro to the Rumen Anaerobe Butyrivibrio fibrisolvens

2001 ◽  
Vol 45 (11) ◽  
pp. 3246-3249 ◽  
Author(s):  
Claire M. Melville ◽  
Karen P. Scott ◽  
Derry K. Mercer ◽  
Harry J. Flint
Keyword(s):  
Resistance Gene ◽  
Pcr Amplification ◽  
Tetracycline Resistance ◽  
Amino Acid Identity ◽  
Tetracycline Resistance Gene ◽  
Butyrivibrio Fibrisolvens ◽  
Acid Identity ◽  
High Level ◽  
Porcine Feces

ABSTRACT A novel tetracycline resistance gene, designatedtet(32), which confers a high level of tetracycline resistance, was identified in the Clostridium-related human colonic anaerobe K10, which also carries tet(W).tet(32) was transmissible in vitro to the rumen anaerobeButyrivibrio fibrisolvens2221R. The predicted gene product oftet(32) has 76% amino acid identity with Tet(O). PCR amplification indicated that tet(32) is widely distributed in the ovine rumen and in porcine feces.

Genetic diversity and relationships among Secale L. based on RAMP markers

2004 ◽  
Vol 1 (2) ◽  
pp. 73-78 ◽  
Author(s):  
Shang Hai-Ying ◽  
Zheng You-Liang ◽  
Wei Yu-Ming ◽  
Wu Wei ◽  
Yan Ze-Hong
Keyword(s):  
Genetic Diversity ◽  
Genetic Relationships ◽  
Pcr Amplification ◽  
Group Method ◽  
Transitional Region ◽  
Arithmetic Average ◽  
Pair Group ◽  
Broad Leaf ◽  
Polymorphic Bands ◽  
High Level

AbstractGenetic diversity and relationships among 21 accessions of Secale L., including three species and 10 subspecies, were evaluated using RAMP markers. Forty-one out of 80 (50.5%) RAMP primers, which produced clear and polymorphic bands, were selected for PCR amplification of genomic DNA. A total of 446 bands were amplified from the 41 primers, and 428 of these bands (about 96%) were polymorphic. Three to 19 polymorphic bands could be amplified from each primer, with an average of 10.4 bands. The RAMP-based genetic similarity (GS) values among the 21 Secale accessions ranged from 0.266 to 0.658, with a mean of 0.449. A high level of genetic variation was found between or within the wild populations and the cultivars. Based on the GS matrix, a dendrogram was constructed using the unweighted pair group method with arithmetic average (UPGMA). All 21 accessions could be distinguished by RAMP markers. Clustering results showed that the genetic diversity of Secale based on RAMP markers was correlated with geographical distribution. Six rye cultivars, originating from Poland, Portugal, Mexico, Hungary, Armenia and Ukraine, were clustered into one group. The six countries are all located in the transitional region of broad-leaf forests between maritime and continental temperate zones, with narrow latitude span. In comparison, the other five cultivars from countries scattered over a region with large latitude span were distributed within different groups or subgroups. Genetic relationships based on RAMP markers had great deviation from the original taxonomy. Some subspecies of the same species were distributed within different groups, while some accessions of different species were closely clustered into one subgroup. These results suggest that RAMP markers could be an effective technique for detecting genetic diversity among Secale and give some useful information about its phylogenic relationships.

Sign in / Sign up

Export Citation Format

Share Document