scholarly journals An Efficient DNA Extraction Method for Polymerase Chain Reaction–Based Detection ofMycobacterium AviumSubspeciesParatuberculosisin Bovine Fecal Samples

2011 ◽  
Vol 23 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Michael Z. Zhang ◽  
Shuping Zhang
1995 ◽  
Vol 27 (3) ◽  
pp. 207-213 ◽  
Author(s):  
Daniele Armaleo ◽  
Philippe Clerc

AbstractA simple DNA extraction method is described, applicable to many different kinds of lichens. The method involves the use of the detergents DTAB and CTAB and yields DNA that can be directly amplified with the polymerase chain reaction or digested with restriction enzymes


2011 ◽  
Vol 23 (6) ◽  
pp. 1189-1196 ◽  
Author(s):  
Silvia Faccini ◽  
Arrigo D. Nigrelli ◽  
Giuliana Franzini ◽  
Carlo Rosignoli ◽  
Ilaria Barbieri ◽  
...  

Quantitative real-time polymerase chain reaction (PCR) has become an important tool for Porcine circovirus-2 (PCV-2) research and diagnosis. However, significant differences in detection limit and quantification data, among laboratories and quantitative real-time PCR methods, have been demonstrated. New efforts are required for providing more accurate and comparable results. The current study is an evaluation of the effects of DNA extraction procedures on PCV-2 quantification in lymph node samples. Differences, greater than 1 log10 copies/g, were shown among PCV-2 loads detected after different extraction procedures. The work highlighted the critical role of the DNA extraction method in PCV-2 quantification by quantitative real-time PCR. This important aspect should be evaluated when comparing data from different laboratories or different studies. The PCV-2 quantification data should not be considered comparable before demonstrating the equivalence of the DNA extraction methods performed.


Author(s):  
Kamaliah Kamaliah

DNA extract is the first phase of molecular research which really influences the quality of DNA isolate. The aim of this study was to find out the comparison of DNA extract by using Phenol-Chloroform and KIT extraction. The visualization of DNA isolate resulted from electrophoresis technique after leptin gen of Aceh’s cow and Madura’s cow were amplificated through in vitro (Polymerase Chain Reaction). There were not any techniques of DNA extraction which can provide excellence results as a whole. Phenol-Chloroform extraction method gives advantage for lower price, more sensitive in isolating DNA, and clear DNA ribbon. Meanwhile, KIT extraction method is easier, fast, and thick DNA produced.


Author(s):  
Mohamed Sabri Esa ◽  
Nur Huda Faujan ◽  
Haitham Abdullah Rajab ◽  
Maryam Mohamed Rehan ◽  
Norlelawati Arifin ◽  
...  

The efficiency of DNA extraction from whole blood using appropriate method is very important for molecular analysis. Therefore, the aim of this study was to compare the purity and concentration of DNA extraction method from bovine (Bos taurus), chicken (Gallus gallus), and porcine (Sus scrofa) blood. The DNA of blood samples was extracted using three types of kit, namely InnuPREP Blood DNA Mini Kit, Wizard Genomic DNA Purification Kit, and QIAamp DNA Blood Mini Kit. The results showed that blood DNA extracted using QIAamp DNA Blood Mini Kit was found to be the most effective and consistently produced high concentrated and pure DNA for three animal samples. The purity of DNA ranged from 1.73 ± 0.05 Å to 1.94 ± 0.21 Å and the range of blood DNA concentration extracted using the QIAamp DNA were between 13.73 ± 2.11 and 25.01 ± 2.08 ng/?l. However, the blood DNA of porcine was not successfully extracted using InnuPREP Blood DNA Mini Kit and Wizard® Genomic DNA Purification Kit. These results were very crucial for the subsequent use of amplification using polymerase chain reaction (PCR) and to facilitate accurate detection in further analysis.


2008 ◽  
Vol 3 (1) ◽  
pp. e29-e29
Author(s):  
B. Sareyyüpoğlu ◽  
A Çelik Ok ◽  
Z. Cantekin ◽  
H. Yardimci ◽  
M. Akan ◽  
...  

1993 ◽  
Vol 5 (3) ◽  
pp. 378-385 ◽  
Author(s):  
Gregory G. Stone ◽  
M. M. Chengappa ◽  
Richard D. Oberst ◽  
Nathan H. Gabbert ◽  
Scott McVey ◽  
...  

The polymerase chain reaction was employed to correlate Salmonella serovars isolated from fecal material of greyhounds suffering from gastroenteritis with those isolated from the diet fed to the greyhounds prior to onset of diarrhea. Kennels around the Abilene, Kansas, area were contacted and supplied with materials needed to collect a portion of the diet each day. With t e onset of diarrhea, the kennels were instructed to ship the fecal material and diet from the previous 10 days to the laboratory for testing. Forty-one fecal samples and corresponding diets were screened for Salmonella, Clostridium perfringens, Campylobacterjejuni, Staphylococcus aureus, Staphylococcus intermedius, and pathogenic (piliated) Escherichia coli by direct culture using standard procedures. The fecal material was also screened for coronavirus and parvovirus using electron microscopy. Thirty-five “normal” fecal samples were screened for all of the above mentioned microorganisms as a control. In addition, the fecal material was screened for E. coli verotoxins I and II and clostridial enterotoxins. A total of 61 Salmonella isolates were recovered from the 41 samples of feces and diet submitted for testing; 31 were recovered from the feces and 30 from the diet. Four Salmonella isolates were recovered from the normal fecal samples. Results obtained by PCR, plasmid profiles, antigenic analysis, and antibiogram profiles indicated that 16 of the 31 isolates recovered from the fecal material were the same strain as that recovered from the diet.


Author(s):  
Dwiyitno Dwiyitno ◽  
Stefan Hoffman ◽  
Koen Parmentier ◽  
Chris Van Keer

Fish and seafood products has been commonly targeted for fraudulent activities. For that reason, authentication of fish and seafood products is important to protect consumers from fraudulent and adulteration practices, as well as to implement traceability regulation. From the viewpoint of food safety, authenticity is beneficial to protect public from serious food poisoning incidents, such as due to ingestion of toxic species. Since DNA based identification depends on the nucleic acid polymerase chain reaction (PCR), the quantity and quality/purity of DNA will contribute significantly to the species authentication. In the present study, different DNA extraction and purification methods (3 classical methods and one commercial kit) were compared to produce the better isolated DNA for PCR amplification. Additionally, different methods for the estimation of DNA concentration and purity which is essential for PCR amplification efficiency were also evaluated. The result showed that classical DNA extraction methods (based on TNES-Urea) yielded a higher amount of DNA (11.30-323.60 ng/g tissue) in comparison to commercial kit/Wizard Promega (5.70-83.45 ng/g tissue). Based on the purity of DNA extract (A260/280), classical DNA extraction method produced relatively similar on DNA quality to the commercial kit (1.79-2.12). Interestingly, all classical methods produced DNA with A260/280 ratio of more than 2.00 on the blue mussel, in contrast with commercial kit. The commercial kit also produced better quality of DNA compared to the classical methods, showing the higher efficiency in PCR amplification. NanoDrop is promising as cheap, robust and safe UV-spectrophotometer method for DNA quantification, as well as the purity evaluation.Keywords: seafood authenticity, DNA isolation, polymerase chain reaction, NanoDrop, Picogreen


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