Detection of Escherichia Coli Using PCR Analysis Without DNA Extraction

This study aimed to detect Escherichia coli directly without DNA extraction. The nucleus membrane and cell membranes of the Escherichia coli are composed of a phospholipid bilayer, damaged if heated at 950C. Pre-denaturation and denaturation of PCR were carried out at 950C. The two stages are thought to break down the Escherichia coli cells, so that the DNA that comes out of the cells can directly become a template in the PCR analysis. In this study, PCR analysis was carried out using Escherichia coli culture, Escherichia coli bacteria culture incubated at 950C, and Escherichia coli bacteria cultures incubated at 650C + on ice as templates. The results showed that PCR analysis using Escherichia coli culture directly and Escherichia coli culture incubated at 650C + on ice as templates produced very thin DNA bands with a size of 580 bp. while PCR analysis using Escherichia coli bacteria culture incubated at 950C as a template produced thick DNA bands with a size of 580 bp. This study's results are very useful for saving time and costs in the detection of Escherichia coli bacteria. The sample to be tested does not need DNA isolation as usual, but only needs to be incubated at 950C for 10 minutes.

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Determining the optimal method for DNA isolation from fruit jams

Czech Journal of Food Sciences ◽

10.17221/340/2017-cjfs ◽

2018 ◽

Vol 36 (No. 2) ◽

pp. 126-132

Author(s):

Sovová Tereza ◽

Křížová Barbora ◽

Ovesná Jaroslava

Keyword(s):

Dna Extraction ◽

Dna Isolation ◽

Extraction Methods ◽

Food Samples ◽

Pcr Analysis ◽

Uv Spectrophotometry ◽

Pcr Assay ◽

Optimal Method ◽

Ctab Method ◽

Dna Extraction Methods

DNA extraction is a crucial step in PCR analysis especially when analysing food samples that can be degraded and can potentially contain PCR-inhibiting substances. In this study, we compared the suitability of three DNA extraction methods – two kits: DNeasy® Plant Mini Kit and NucleoSpin® Food, and the CTAB method – for DNA extraction from commercial fruit jams. Fourteen jams with different contents of fruit, sugar and other additives were extracted in triplicate using the above-mentioned methods directly and after a washing step. The concentration and optical density were analysed using UV spectrophotometry and the amplifiability of the obtained DNA was evaluated using a PCR assay targeting a sequence coding for chloroplast tRNA-Leu. Samples isolated using the NucleoSpin® Food kit contained non-amplifiable DNA in eight cases, and samples isolated using the CTAB method could not be quantified. The DNeasy® Plant Mini Kit thus proved to be the most suitable method, since well-amplifiable DNA was obtained for all the analysed samples.

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Effect of Meat Processing and Cooking on DNA Extraction and Detection of Meat Adulteration in Mutton Rista (Kashmiri Meat Product)

The Indian Journal of Nutrition and Dietetics ◽

10.21048/ijnd.2016.53.4.8403 ◽

2016 ◽

Vol 53 (4) ◽

pp. 459

Author(s):

Mohammad Mansoor Bhat ◽

Heena Jalal ◽

Parveez Ahmad Para ◽

Subha Ganguly

Keyword(s):

Multiplex Pcr ◽

Dna Extraction ◽

Dna Isolation ◽

Meat Product ◽

Pcr Analysis ◽

Meat Processing ◽

Buffalo Meat ◽

Study Material ◽

Dna Quality ◽

Meat Samples

The processing and cooking of meat, during meat product preparation, affects the DNA quality and its concentration during DNA isolation. In this study, the effect of processing and cooking, during Rista preparation, on meat speciation of beef and buffalo meat in mutton Rista was studied. The study material involved three types of meat i.e. unprocessed meat, Rista emulsion and the final cooked Rista product. In each type of meat, pure meat samples of mutton, beef and buffalo meat were studied along with the adulterated mutton sample having 10% beef and 10% buffalo meat adulteration level. The meat samples were subjected to mtDNA isolation and multiplex PCR analysis. The results of this study showed that processing and cooking decreases the concentration of extracted DNAs but does not affect the detection of beef and buffalo meat in adulterated mutton Rista (unprocessed, processed and cooked) at 10% level of adulteration.

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Faculty Opinions recommendation of Toxicological impact studies based on Escherichia coli bacteria in ultrafine ZnO nanoparticles colloidal medium.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1012510.372000 ◽

2006 ◽

Author(s):

Anthony Czarnik

Keyword(s):

Escherichia Coli ◽

Zno Nanoparticles ◽

Impact Studies ◽

Escherichia Coli Bacteria

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Faculty Opinions recommendation of RNA dynamics in live Escherichia coli cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1021598.246622 ◽

2004 ◽

Author(s):

Tina Henkin

Keyword(s):

Escherichia Coli ◽

Rna Dynamics ◽

Escherichia Coli Cells

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Faculty Opinions recommendation of Deletion of penicillin-binding protein 5 (PBP5) sensitises Escherichia coli cells to beta-lactam agents.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1649962.1152060 ◽

2010 ◽

Author(s):

Samuel Kariuki

Keyword(s):

Escherichia Coli ◽

Binding Protein ◽

Penicillin Binding Protein ◽

Beta Lactam ◽

Escherichia Coli Cells

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Extraction of DNA from a Wide Range of Objects by Treatment with Ammonium Salts

Biotekhnologiya ◽

10.21519/0234-2758-2020-36-6-98-106 ◽

2020 ◽

Vol 36 (6) ◽

pp. 98-106

Author(s):

E.I. Levitin ◽

B.V. Sviridov ◽

O.V. Piksasova ◽

T.E. Shustikova

Keyword(s):

Dna Extraction ◽

Dna Isolation ◽

Tissue Samples ◽

New Approach ◽

Cell Wall Disruption ◽

Wide Range ◽

Low Salt ◽

Mammalian Blood ◽

Plasmid Extraction ◽

Two Hybrid

Currently, simple, rapid, and efficient techniques for DNA isolation from a wide range of organisms are in demand in biotechnology and bioinformatics. A key (and often limiting) step is the cell wall disruption and subsequent DNA extraction from the disintegrated cells. We have developed a new approach to DNA isolation from organisms with robust cell walls. The protocol includes the following steps: treatment of cells or tissue samples with ammonium acetate followed by cell lysis in low-salt buffer with the addition of SDS. Further DNA extraction is carried out according to standard methods. This approach is efficient for high-molecular native DNA isolation from bacteria, ascomycetes, yeast, and mammalian blood; it is also useful for express analysis of environmental microbial isolates and for plasmid extraction for two-hybrid library screening. express method for DNA isolation; ammonium salt treatment (в русских ключевых такой порядок), osmotic breakage of cells This study was financially supported by the NRC "Kurchatov Institute"-GOSNIIGENETIKA Kurchatov Genomic Center.

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