Chelex DNA extraction Protocol v1

Author(s):  
Sarah Chang ◽  
Michael Russello

Laboratory protocol for DNA extraction from animal tissue with Chelex.

2018 ◽  
Vol 24 (2) ◽  
pp. 77-86
Author(s):  
Sri Wening ◽  
Agus Eko Prasetyo ◽  
Tjut Ahmad Perdana Rozziansha ◽  
Agus Susanto

African pollination weevil (Elaeidobius kamerunicus Faust) has an important role in the productivity of Indonesian oil palm plantation. Up to now, there has not been a comprehensive biological study of the species at molecular level. The basic knowledge is very useful for exploitation of the weevil for effective oil palm fruit set development. This research aimed to obtain DNA extraction protocol of E. kamerunicus for DNA fingerprinting of the species. Results showed that using a DNA extraction kit,material disruption by using micro pestle resulted the highest quantity of DNA, while there were no significant differences of resulted DNA quantity among treatments using tissue lyser for material disruption. DNA extracted by using micro pestle or tissue lyser for material disruption is adequate for DNA fingerprinting using AFLP (Amplified Fragment Length Polymorphism) and sequencing techniques.


2011 ◽  
Vol 39 (1) ◽  
pp. 236-242 ◽  
Author(s):  
S.N. Sharma ◽  
V. Kumar ◽  
G. Singh ◽  
R. Sharma

2013 ◽  
Vol 12 (1) ◽  
pp. 282-292 ◽  
Author(s):  
B. Canto-Canché ◽  
M. Tzec-Simá ◽  
J.I. Vázquez-Loría ◽  
H. Espadas-Álvarez ◽  
B.H. Chí-Manzanero ◽  
...  

Author(s):  
Priscila Lie Tobouti ◽  
Juliana Seo ◽  
Michella Bezerra Lima ◽  
Bruno Tavares Sedassari ◽  
Norberto Nobuo Sugaya ◽  
...  

<p><strong>Objective: </strong>To compare the diagnostic accuracy of immunohistochemistry compared to real-time PCR (using a simple phenol-chloroform DNA extraction protocol) in the detection of HHV8 in small oral biopsies of Kaposi sarcoma. Also to validate the use of this DNA extraction protocol to extract HHV8 DNA.</p><p><strong>Material and methods:</strong> Seventeen cases of oral KS were examined. Data including gender, age, and anatomic location were obtained from patient´s records and histological sections stained with hematoxylin and eosin (H&amp;E) were reviewed. Immunohistochemistry was used to detect HHV8 in lesions of interest, as well as real-time PCR.</p><p><strong>Results: </strong>All the patients were HIV positive, the mean age was 35.5 years old, and the affected oral sites were palate (47%), gingiva (29.4%), tongue (11.8%), lip (5.9%), and oral floor (5.9%). Fifteen samples showed positive staining for HHV8 and only two samples were negative. The same results were observed using real-time PCR HHV8-DNA detection.</p><p><strong>Relevance: </strong>Our findings suggest that immunohistochemistry is faster and cheaper to perform than real-time PCR and was shown to have similar levels of sensitivity and accuracy for detection of HHV8 even in small biopsies. Additionally DNA extraction using a non-commercial kit, as done in this study can further reduce the costs to a pathology service.</p>


2011 ◽  
Vol 56 (5) ◽  
pp. 1235-1240 ◽  
Author(s):  
Julieta Kopka ◽  
Monika Leder ◽  
Stella M. Jaureguiberry ◽  
Gottfried Brem ◽  
Gabriel O. Boselli

2019 ◽  
Vol 12 (9) ◽  
pp. 1998-2008
Author(s):  
Xiong Xiong ◽  
Manhong Huang ◽  
Fangying Yuan ◽  
Lixia Lu ◽  
Xiaohui Xiong

2020 ◽  
Vol 8 (4) ◽  
pp. 532 ◽  
Author(s):  
Victor Guerra ◽  
Lukas Beule ◽  
Ena Lehtsaar ◽  
Hui-Ling Liao ◽  
Petr Karlovsky

As our understanding of soil biology deepens, there is a growing demand for investigations addressing microbial processes in the earth beneath the topsoil layer, called subsoil. High clay content in subsoils often hinders the recovery of sufficient quantities of DNA as clay particles bind nucleic acids. Here, an efficient and reproducible DNA extraction method for 200 mg dried soil based on sodium dodecyl sulfate (SDS) lysis in the presence of phosphate buffer has been developed. The extraction protocol was optimized by quantifying bacterial 16S and fungal 18S rRNA genes amplified from extracts obtained by different combinations of lysis methods and phosphate buffer washes. The combination of one minute of bead beating, followed by ten min incubation at 65°C in the presence of 1 M phosphate buffer with 0.5% SDS, was found to produce the best results. The optimized protocol was compared with a commonly used cetyltrimethylammonium bromide (CTAB) method, using Phaeozem soil collected from 60 cm depth at a conventional agricultural field and validated on five subsoils. The reproducibility and robustness of the protocol was corroborated by an interlaboratory comparison. The DNA extraction protocol offers a reproducible and cost-effective tool for DNA-based studies of subsoil biology.


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