scholarly journals OPTIMASI METODE LISIS ALKALI UNTUK MENINGKATKAN KONSENTRASI PLASMID

2015 ◽  
Vol 2 (2) ◽  
pp. 60
Author(s):  
Dudi Hardianto ◽  
Alfik Indarto ◽  
Nurtjahjo Dwi Sasongko
Keyword(s):  
Escherichia Coli ◽  
Molecular Biology ◽  
Low Cost ◽  
Cesium Chloride ◽  
Alkaline Lysis ◽  
Isolation Methods ◽  
Plasmid Isolation ◽  
Biology Laboratory

Plasmids are extra chromosomal molecules of DNA that replicate autonomously and found in prokaryote and eukaryote cells. There are a number of methods that are used to isolate plasmids, such as alkaline lysis, boiling lysis, using cesium chloride, and chromatography. Amongst the disadvantages in plasmid isolation methods are lengthy time especially when handling a large number of samples, high cost, and low purity. Alkaline lysis is the most popular for plasmid isolation because of its simplicity, relatively low cost, and reproducibility. This method can be accomplished in 50 minutes to one hour. In this research, the alkaline lysis method was developed to obtain suitable plasmid for applications in a molecular biology laboratory. The aim of this research was to reduce contaminants and improve yield of plasmid. The result of isolation of pICZA plasmid in Escherichia coli gave the concentration of 3.3 to 3.8 µg/µL with the purity of 1.99.Keywords: Plasmid isolation, pICZ A, Escherichia coli, rapid, alkaline lysis  ABSTRAKPlasmid merupakan molekul DNA ekstrakromosomal yang bereplikasi secara mandiri dan ditemukan dalam sel prokariot dan eukariot. Banyak metode yang digunakan untuk isolasi plasmid, seperti: lisis alkali, lisis dengan pemanasan, penggunaan sesium klorida, dan kromatografi. Kelemahan beberapa metode isolasi DNA adalah waktu isolasi yang lama terutama saat isolasi plasmid dalam jumlah banyak, mahal dan kemurniannya yang rendah. Metode lisis alkali merupakan metode yang sangat umum untuk isolasi plasmid karena mudah dilakukan, relatif murah, dan reprodusibilitas. Metode ini dapat dilakukan dalam 50 menit sampai 1 jam. Pada penelitian ini dikembangkan metode lisis alkali untuk memperoleh plasmid yang sesuai untuk penggunaan di laboratorium biologi molekuler. Tujuan dari penelitian ini adalah untuk mengurangi jumlah kontaminan dan meningkatkan konsentrasi plasmid. Hasil isolasi plasmid pICZA dalam Escherichia coli mempunyai konsentrasi antara 3,3 sampai 3,8 µg/µL dan kemurniannya 1,99.Kata Kunci: Isolasi plasmid, pICZ A, Escherichia coli, cepat, lisis alkali

Low Cost Controlled Composting of Refuse and Sewage Sludge

1987 ◽  
Vol 19 (5-6) ◽  
pp. 839-845 ◽  
Author(s):  
J. T. Pereira-Neto ◽  
E. I. Stentiford ◽  
D. D. Mara
Keyword(s):  
Escherichia Coli ◽  
Sewage Sludge ◽  
Low Cost ◽  
Control Methods ◽  
Fixed Rate ◽  
The Past ◽  
Temperature Feedback ◽  
Forced Aeration ◽  
Domestic Refuse ◽  
Faecal Streptococci

The forced aeration static pile composting system was used to compost mixtures of domestic refuse and sewage sludge. Several different control methods have been evaluated over the past four years from simple, low cost fixed rate aeration timers to microcomputer based systems. Their relative merits are considered. In a compost pile using temperature feedback control the number of Escherichia coli were reduced from 107 org./g to less than 102 org./g. within 16 days. Faecal streptococci were reduced from 107 to less than 102 org./g within 30 days. The process consistently produced a good quality sanitised material under a range of control regimes.

scholarly journals Horizontal Transfer of Antimicrobial Resistance by Extended-Spectrum β Lactamase-Producing Enterobacteriaceae

2011 ◽  
Vol 3 (01) ◽  
pp. 037-042 ◽  
Author(s):  
Varsha K Vaidya
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Resistance Genes ◽  
Environmental Conditions ◽  
Horizontal Transfer ◽  
Hospital Setting ◽  
Alkaline Lysis ◽  
Antibiotic Resistant ◽  
Extended Spectrum ◽  
Synergy Test

ABSTRACT Background: The purpose of this work was to study the acquisition of new antibiotic-resistant genes carried by extended spectrum β-lactamase (ESBL)-producing Enterobacteriaceae via horizontal transfer to understand their rampant spread in the hospitals and in the community. Materials and Methods: A retrospective analysis of 120 ESBL screen-positive isolates of Escherichia coli and Klebsiella pneumoniae, which were subjected to antimicrobial susceptibility testing, was carried out. The Double Disc Synergy Test (DDST) and Inhibitor-Potentiation Disc Diffusion Test (IPDD) were employed for confirmation of ESBL activity. The transferability of the associated antibiotic resistance for amoxicillin, amikacin, gentamicin, cefotaxime and ceftriaxone was elucidated by intra- and intergenus conjugation in Escherichia coli under laboratory as well as under simulated environmental conditions. Transformation experiments using plasmids isolated by alkaline lysis method were performed to study the transferability of resistance genes in Klebsiella pneumoniae isolates. Results : ESBL production was indicated in 20% each of the Escherichia coli and Klebsiella pneumoniae isolates. All the ESBL isolates showed co- resistance to various other groups of antibiotics, including 3GC antibiotics, though all the isolates were sensitive to both the carbapenems tested. Conjugation-mediated transfer of resistance under laboratory as well as environmental conditions at a frequency of 3-4 x 10-5 , and transformation-mediated dissemination of cefotaxime and gentamicin resistance shed light on the propensity of ESBL producers for horizontal transfer. Conclusions: The transfer of resistant markers indicated availability of a large pool of resistance genes in the hospital setting as well as in the environment, facilitating long-term persistence of organisms.

scholarly journals Rapid, simple, low-cost smartphone-based fluorescence detection of Escherichia coli

2021 ◽  
Vol 14 (3) ◽  
pp. 167-174
Author(s):  
Dante Rojas-Barboza ◽  
◽  
Edward Park ◽  
Rolfe Sassenfeld ◽  
Jeremy Winder ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fluorescence Detection ◽  
Low Cost
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