Toward Metrological Traceability for DNA Fragment Ratios in GM Quantification. 1. Effect of DNA Extraction Methods on the Quantitative Determination of Bt176 Corn by Real-Time PCR

2007 ◽  
Vol 55 (9) ◽  
pp. 3249-3257 ◽  
Author(s):  
Philippe Corbisier ◽  
Wim Broothaerts ◽  
Sabrina Gioria ◽  
Heinz Schimmel ◽  
Malcolm Burns ◽  
...  
Keyword(s):  
Quantitative Determination ◽  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Metrological Traceability ◽  
Extraction Methods ◽  
Dna Fragment ◽  
Dna Extraction Methods

scholarly journals Evaluation of DNA extraction methods for quantifying Helicobacter pylori in water with PCR inhibitors

2021 ◽  
Author(s):  
Eun-Sook Lee ◽  
So-Yang Cha ◽  
Jong-Soon Jung
Keyword(s):  
Helicobacter Pylori ◽  
Real Time ◽  
Dna Extraction ◽  
Water Samples ◽  
Real Time Pcr ◽  
Heavy Rain ◽  
Extraction Methods ◽  
Pcr Inhibitors ◽  
H Pylori ◽  
Dna Extraction Methods

Abstract DNA extraction methods were evaluated to reduce PCR inhibitors and quantify Helicobacter pylori directly from water samples using real-time PCR. Three nucleic acid extraction methods were evaluated for different types of water samples. While the QIAamp DNA mini kit for tissue was suitable for DNA extraction from treated water, the QIAamp DNA stool mini kit was still efficient in analyzing samples from river water after heavy rain and with high concentration of PCR inhibitors. The FastDNA SPIN Kit for Soil could extract DNA effectively from microbes in river and stream waters without heavy rain. Immunomagnetic separation (IMS) was used prior to DNA extraction and was a useful tool for reducing PCR inhibitors in influent and stream samples. H. pylori in various waters could be quantified directly by real-time PCR while minimizing the effect of PCR inhibitors by an appropriate method through the evaluation of DNA extraction methods considering the characteristics of the matrix water. The findings of the present study suggest that the types or characteristics of water sample by source and precipitation are an important factor in detecting H. pylori and they can be applied when detecting and monitoring of other pathogens in water.

A comparative study of DNA extraction methods to be used in real-time PCR based quantification of ochratoxin A-producing molds in food products

Food Control ◽  
2012 ◽  
Vol 25 (2) ◽  
pp. 666-672 ◽  
Author(s):  
Alicia Rodríguez ◽  
Mar Rodríguez ◽  
M. Isabel Luque ◽  
Annemarie F. Justesen ◽  
Juan J. Córdoba
Keyword(s):  
Comparative Study ◽  
Real Time ◽  
Ochratoxin A ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Food Products ◽  
Extraction Methods ◽  
Dna Extraction Methods

scholarly journals Comparison of DNA extraction methods for Aspergillus fumigatus using real-time PCR

2006 ◽  
Vol 55 (9) ◽  
pp. 1187-1191 ◽  
Author(s):  
Lisa J. Griffiths ◽  
Martin Anyim ◽  
Sarah R. Doffman ◽  
Mark Wilks ◽  
Michael R. Millar ◽  
...  
Keyword(s):  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Diagnostic Technique ◽  
Extraction Methods ◽  
Simple Method ◽  
Bead Beating ◽  
Fungal Dna ◽  
Dna Extraction Methods ◽  
High Level

Newer methods such as PCR are being investigated in order to improve the diagnosis of invasive aspergillosis. One of the major obstacles to using PCR to diagnose aspergillosis is a reliable, simple method for extraction of the fungal DNA. The presence of a complex, sturdy cell wall that is resistant to lysis impairs extraction of the DNA by conventional methods employed for bacteria. Numerous fungal DNA extraction protocols have been described in the literature. However, these methods are time-consuming, require a high level of skill and may not be suitable for use as a routine diagnostic technique. Here, a number of extraction methods were compared: a freeze–thaw method, a freeze–boil method, enzyme extraction and a bead-beating method using Mini-BeadBeater-8. The quality and quantity of the DNA extracted was compared using real-time PCR. It was found that the use of a bead-beating method followed by extraction with AL buffer (Qiagen) was the most successful extraction technique, giving the greatest yield of DNA, and was also the least time-consuming method assessed.

Comparison of DNA extraction methods used to detect bacterial and yeast DNA from spiked whole blood by real-time PCR

2017 ◽  
Vol 140 ◽  
pp. 61-66 ◽  
Author(s):  
Libera M. Dalla-Costa ◽  
Luis G. Morello ◽  
Danieli Conte ◽  
Luciane A. Pereira ◽  
Jussara K. Palmeiro ◽  
...  
Keyword(s):  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Whole Blood ◽  
Extraction Methods ◽  
Dna Extraction Methods
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