An Efficient DNA Extraction Protocol for Successful PCR Detection of Banana bunchy top virus from Banana Leaves

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.

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A modified DNA extraction protocol and its utility in seed genetic purity assessment

Seed Science and Technology ◽

10.15258/sst.2011.39.1.24 ◽

2011 ◽

Vol 39 (1) ◽

pp. 236-242 ◽

Cited By ~ 3

Author(s):

S.N. Sharma ◽

V. Kumar ◽

G. Singh ◽

R. Sharma

Keyword(s):

Dna Extraction ◽

Genetic Purity ◽

Purity Assessment ◽

Extraction Protocol ◽

Modified Dna

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Improving PCR detection of prey in molecular diet studies: importance of group‐specific primer set selection and extraction protocol performances

Molecular Ecology Resources ◽

10.1111/1755-0998.12029 ◽

2012 ◽

Vol 13 (1) ◽

pp. 117-127 ◽

Cited By ~ 27

Author(s):

Diane Zarzoso‐Lacoste ◽

Emmanuel Corse ◽

Eric Vidal

Keyword(s):

Pcr Detection ◽

Specific Primer ◽

Extraction Protocol ◽

Group Specific Primer

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Small volume fungal genomic DNA extraction protocol for Illumina genome v2 (protocols.io.3w9gph6)

protocols.io ◽

10.17504/protocols.io.3w9gph6 ◽

2019 ◽

Author(s):

Jana M ◽

Lilly Moore

Keyword(s):

Dna Extraction ◽

Genomic Dna ◽

Small Volume ◽

Genomic Dna Extraction ◽

Extraction Protocol

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Methodology Simple and inexpensive DNA extraction protocol for studying the bacterial composition of sludges used in microbial fuel cells

Genetics and Molecular Research ◽

10.4238/2013.february.4.2 ◽

2013 ◽

Vol 12 (1) ◽

pp. 282-292 ◽

Cited By ~ 3

Author(s):

B. Canto-Canché ◽

M. Tzec-Simá ◽

J.I. Vázquez-Loría ◽

H. Espadas-Álvarez ◽

B.H. Chí-Manzanero ◽

...

Keyword(s):

Fuel Cells ◽

Dna Extraction ◽

Microbial Fuel Cells ◽

Bacterial Composition ◽

Extraction Protocol

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Comparison of Four Commercial DNA Extraction Kits for PCR Detection of Listeria monocytogenes, Salmonella, Escherichia coli O157:H7, and Staphylococcus aureus in Fresh, Minimally Processed Vegetables

Journal of Food Protection ◽

10.4315/0362-028x-71.10.2110 ◽

2008 ◽

Vol 71 (10) ◽

pp. 2110-2114 ◽

Cited By ~ 25

Author(s):

P. ELIZAQUÍVEL ◽

R. AZNAR

Keyword(s):

Escherichia Coli ◽

Staphylococcus Aureus ◽

Listeria Monocytogenes ◽

Dna Extraction ◽

Pcr Detection ◽

Escherichia Coli O157 ◽

Minimally Processed ◽

Green Pepper ◽

Minimally Processed Vegetables ◽

And Staphylococcus Aureus

Four commercial DNA extraction methods, PrepMan Ultra (Applied Biosystems), InstaGene Matrix (BioRad), DNeasy Tissue kit (Qiagen), and UltraClean (MoBio), were tested for PCR detection of Listeria monocytogenes, Escherichia coli O157: H7, Salmonella, and Staphylococcus aureus in fresh, minimally processed vegetables. For comparative purposes, sensitivity assays with specific PCRs were carried out after DNA extraction with the four methods in green pepper, broccoli, and onion artificially inoculated with the four pathogens separately. As confirmed by statistical analysis, the DNeasy Tissue kit rendered the highest sensitivity values in the three matrices assayed for Salmonella, L. monocytogenes, and E. coli O157:H7 and in onion for S. aureus. Despite being the most expensive of the methods compared, the DNeasy Tissue Kit can be successfully applied for any of the four most commonly studied pathogens, thus saving time and overall reducing the cost of the analysis.

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ORAL HIV-ASSOCIATED KAPOSI SARCOMA: A COMPARISON BETWEEN IMMUNOHISTOCHEMISTRY AND qPCR TECHNIQUES FOR DETECTION OF HHV8

Clinical and Laboratorial Research in Dentistry ◽

10.11606/issn.2357-8041.clrd.2015.97224 ◽

2015 ◽

Vol 21 (1) ◽

pp. 29

Author(s):

Priscila Lie Tobouti ◽

Juliana Seo ◽

Michella Bezerra Lima ◽

Bruno Tavares Sedassari ◽

Norberto Nobuo Sugaya ◽

...

Keyword(s):

Real Time ◽

Dna Extraction ◽

Real Time Pcr ◽

Kaposi Sarcoma ◽

Positive Staining ◽

Extraction Protocol ◽

Simple Phenol ◽

Two Samples ◽

Hematoxylin And Eosin ◽

The Mean

Objective: 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.Material and methods: 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&E) were reviewed. Immunohistochemistry was used to detect HHV8 in lesions of interest, as well as real-time PCR.Results: 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.Relevance: 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.

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