Plasmid DNA Extraction from E. coli Using Alkaline Lysis Method

BIO-PROTOCOL ◽  
2011 ◽  
Vol 1 (1) ◽  
Author(s):  
He, F. Fanglian
Keyword(s):  
Dna Extraction ◽  
Plasmid Dna ◽  
Alkaline Lysis ◽  
E Coli

Plasmid DNA extraction v2

2021 ◽  
Author(s):  
Shuning Guo
Keyword(s):  
Dna Extraction ◽  
Plasmid Dna ◽  
E Coli

This protocol is used to extract plasmid DNA from E. coli.

Plasmid DNA extraction v2

2021 ◽  
Author(s):  
Shuning Guo
Keyword(s):  
Dna Extraction ◽  
Plasmid Dna ◽  
E Coli

This protocol is used to extract plasmid DNA from E. coli.

Screening of Metal and Antibiotic Resistance in Beta-lactamase Producing Coliform Bacteria from Hospital Wastewater of Northern India

2020 ◽  
Vol 14 (1) ◽  
pp. 63-77 ◽  
Author(s):  
Manzar Alam ◽  
Mohd Imran ◽  
Syed Sayeed Ahmad
Keyword(s):  
Plasmid Dna ◽  
Metal Tolerance ◽  
Coliform Bacteria ◽  
Diffusion Method ◽  
Hazard Evaluation ◽  
Beta Lactamase ◽  
Hospital Wastewater ◽  
Alkaline Lysis ◽  
Potential Health ◽  
E Coli

Aim: Our exploration work has uncovered the different anti-toxin/metal tolerance and patterns against the heavy metal resistant coliform microscopic organisms from the aquatic waste of the hospital. It might give new routes for the treatment of irresistible ailments particularly by coliform and critical for hazard evaluation as well as hazard management associated with the effluents of the hospital. Background: The higher use of pharmaceuticals, Radionuclides, and other antimicrobial solvents are the major source of metals in hospital wastewater. The hospital aquatic environment has a high content of both organic and inorganic matter with living organisms. Bacteria can resist an antimicrobial agent by producing extracellular enzymes that eliminate antibiotics and metal toxicity. In this study, we covered the existing patent literature in this area. New patents in the areas of topically applied antibiotics and agents that can potentiate the achievement of existing antibiotics may extend their helpful lifetime. Methods: Samples were collected from three different Departments of King George Medical University, Lucknow during the month of December to May (2015-16). Isolation and metal tolerance of coliform isolates were done on metal amended plates. The antibiotic sensitivity test was done by disc diffusion method. The plasmid DNA of bacterial isolates was done by the alkaline lysis method. The conjugation study was also performed in wastewater as well as a nutrient medium. Results: Maximum isolates demonstrated their MICs at 400, 800 and 1600 μg/ml against all the metals, respectively. The high level of resistance was observed against Methicillin (88.32%, 80.60%) followed by penicillin (75%, 76%), Cephradin (59.52%, 28.84%) and least to Gentamycine (1.92%, 5.76) in E. coli and Enterobacter, respectively. Of 70%, 78% E. coli and Enterobacter isolates produce beta-lactamase activity. Six amino acid residues namely, Glu104, Tyr105, Asn132, Asn170, Ala237, and Gly238 of the beta-lactamase were found in the common interaction with the selected drugs. Plasmid DNA size ranged between 48-58.8 kb. The conjugation experiments showed a higher transfer frequency (5.5×10-1 and 3.6×10-1) rate among antibiotics and metals tested. Conclusion: The finding of this study presents a potential health problem as the predominant coliform species have increasingly been associated with outbreaks of hospital infections. It is recommended that hospital waste must be properly treated before its release into the environment.

Plasmid DNA extraction v1

2021 ◽  
Author(s):  
Shuning Guo
Keyword(s):  
Dna Extraction ◽  
Plasmid Dna ◽  
E Coli

This protocol is used to extract plasmid DNA from E. coli.

Automation Protocol for Plasmid DNA Extraction from E. coli v1 (protocols.io.bjjvkkn6)

protocols.io ◽  
2020 ◽  
Author(s):  
Nina Alperovich ◽  
Jane Romantseva ◽  
Olga Vasilyeva ◽  
David Ross
Keyword(s):  
Dna Extraction ◽  
Plasmid Dna ◽  
E Coli

Quality Improvement of the DNA extracted by boiling method in Gram negative bacteria

2017 ◽  
Vol 6 (04) ◽  
pp. 5347 ◽  
Author(s):  
Omar B. Ahmed* ◽  
Anas S. Dablool
Keyword(s):  
Dna Extraction ◽  
Control Method ◽  
Extraction Procedure ◽  
Cost Effective ◽  
High Yield ◽  
Gram Negative Bacteria ◽  
Bacterial Dna ◽  
Gram Negative ◽  
E Coli

Several methods of Deoxyribonucleic acid (DNA) extraction have been applied to extract bacterial DNA. The amount and the quality of the DNA obtained for each one of those methods are variable. The study aimed to evaluate bacterial DNA extraction using conventional boiling method followed by alcohol precipitation. DNA extraction from Gram negative bacilli was extracted and precipitated using boiling method with further precipitation by ethanol. The extraction procedure performed using the boiling method resulted in high DNA yields for both E. coli and K. pneumoniae bacteria in (199.7 and 285.7μg/ml, respectively) which was close to control method (229.3 and 440.3μg/ml). It was concluded that after alcohol precipitation boiling procedure was easy, cost-effective, and applicable for high-yield quality of DNA in Gram-negative bacteria.

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