Generation of chromosomal DNA during alkaline lysis and removal by reverse micellar extraction

Tschapalda, Kirsten; Streitner, Nadine; Voss, Carsten; Flaschel, Erwin

doi:10.1007/s00253-009-2088-3

Cited by 3 publications

(1 citation statement)

References 17 publications

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“…Total genomic DNA of all isolates was extracted using an alkaline lysis extraction method. 19 , 20 Briefly, fungal pellets were resuspended sequentially in Solution I (0.9% w/v glucose, 25 mmol/L Tris, 6 mmol/L ethylenediaminetetraacetic acid, pH 8.0) and Solution II (1% SDS, 0.2 mol/L sodium hydroxide), followed by precipitation with Solution III (12% sodium acetate, 12% acetic acid). The supernatant was collected and successively treated with phenol:chloroform:isoamyl alcohol (25:24:1) and chloroform:isoamyl alcohol (24:1).…”

Section: Methodsmentioning

confidence: 99%

Genetic variation of Sporothrix globosa isolates from diverse geographic and clinical origins in China

Zhao

Cui

et al. 2017

Emerging Microbes & Infections

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Sporothrix globosa is the main causative agent of sporotrichosis, a common mycosis that usually affects the skin, in China. Despite increasing efforts in the molecular identification of this fungal pathogen, its modes of transmission and epidemiology remain poorly understood. The goals of this study were to assess the genetic diversity of S. globosa using amplified fragment length polymorphism (AFLP) analysis and to assess the correlation of AFLP profiles with the geographic origins, growth rates, clinical forms, and antifungal susceptibilities of S. globosa isolates. AFLP analysis of 225 clinical S. globosa isolates from eight provinces or municipalities in China identified eight distinct clustering groups (I–VIII), with groups I, II and IV being the most common. The AFLP genotypes showed distinct distribution patterns among different regions within Jilin Province and between northern and southern China, but there was no obvious association between the AFLP genotypes and the growth rates, clinical forms or antifungal susceptibilities of the S. globosa isolates. These results expand our understanding of the genetic variation of S. globosa and suggest that AFLP analysis is a potentially useful tool for studying the epidemiology of this fungal pathogen.

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Section: Methodsmentioning

confidence: 99%

Genetic variation of Sporothrix globosa isolates from diverse geographic and clinical origins in China

Zhao

Cui

et al. 2017

Emerging Microbes & Infections

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Comparison of Alkaline Lysis with Electroextraction and Optimization of Electric Pulses to Extract Plasmid DNA from Escherichia coli

et al. 2013

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The use of plasmid DNA (pDNA) as a pharmaceutical tool has increased since it represents a safer vector for gene transfer compared to viral vectors. Different pDNA extraction methods have been described; among them is alkaline lysis, currently the most commonly used. Although alkaline lysis represents an established method for isolation of pDNA, some drawbacks are recognized, such as entrapment of pDNA in cell debris, leading to lower pDNA recovery; the time-consuming process; and increase of the volume due to the buffers used, all leading to increased cost of production. We compared the concentration of extracted pDNA when two methods for extracting pDNA from Escherichia coli were used: alkaline lysis and a method based on membrane electroporation, electroextraction. At the same time, we also studied the effect of different pulse protocols on bacterial inactivation. The concentration of pDNA was assayed with anion exchange chromatography. When alkaline lysis was used, two incubations of lysis time (5 and 10 min) were compared in terms of the amount of isolated pDNA. We did not observe any difference in pDNA concentration regardless of incubation time used. In electroextraction, different pulse protocols were used in order to exceed the pDNA concentration obtained by alkaline lysis. We show that electroextraction gives a higher concentration of extracted pDNA than alkaline lysis, suggesting the use of electroporation as a potentially superior method for extracting pDNA from E. coli. In addition, electroextraction represents a quicker alternative to alkaline lysis for extracting pDNA.

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Reverse Micellar Extraction of Bioactive Compounds for Food Products

Hemavathi¹,

Hebbar²,

Raghavarao³

2011

Enhancing Extraction Processes in the Food Industry

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Generation of chromosomal DNA during alkaline lysis and removal by reverse micellar extraction

Cited by 3 publications

References 17 publications

Genetic variation of Sporothrix globosa isolates from diverse geographic and clinical origins in China

Genetic variation of Sporothrix globosa isolates from diverse geographic and clinical origins in China

Comparison of Alkaline Lysis with Electroextraction and Optimization of Electric Pulses to Extract Plasmid DNA from Escherichia coli

Reverse Micellar Extraction of Bioactive Compounds for Food Products

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