Preparation of Genomic <scp>DNA</scp> from Plant Tissue

Richards, Eric J.; Reichardt, Mark; Rogers, Sharon A.

doi:10.1002/0471142727.mb0203s27

Cited by 150 publications

(110 citation statements)

References 6 publications

(11 reference statements)

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“…It is a known fact that contaminants like polysaccharides and polyphenols are, in general, difficult to be removed from the DNA as they often adsorb and co-precipitate with the DNA (Murray and Thompson, 1980) and interfere with PCR and restriction endonuclease digestion. Polysaccharides also interfere with several biological enzymes such as polymerases, ligases and restriction endonucleases (Shioda and Murakami, 1987;Richards, 1988). …”

Section: Resultsmentioning

confidence: 99%

A novel and efficient protocol for the isolation of genomic DNA from<br>mulberry (Morus L.)

Anuradhabr

Vijayanbr

Manjula

2013

Emir. J. Food Agric

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The purity of DNA is one of the major factors affecting the success of Genomic studies. Nucleic acid isolation from polyphenol rich plants fail to produce good quality DNA or RNA as polyphenols adhere and interfere with DNA during isolation. An improvised, simple and inexpensive protocol has been developed for extracting genomic DNA from Mulberry (Morus spp.). The purity of the DNA as revealed by the ratios of absorbance at 260/280 nm (A 260/280) and 260/230 nm (A 260/230) was closer to 2.0. Genomic DNA analyzed for analytical applications like restriction digestion and PCR amplification with molecular markers viz., Inter Simple Sequence Repeats (ISSR), Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeat (SSR) primers further confirmed the purity of the DNA. A modified method of silver staining was employed for the resolution of SSR amplified products. Physiologically mature leaf was found more suitable for getting quality DNA in mulberry.

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Section: Resultsmentioning

confidence: 99%

A novel and efficient protocol for the isolation of genomic DNA from<br>mulberry (Morus L.)

Anuradhabr

Vijayanbr

Manjula

2013

Emir. J. Food Agric

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“…Genomic DNA was isolated using a CTAB (cetyl trimethylammonium bromide) method [32]. The genomic DNA samples of the control plants and transformed T0 lines were analyzed by PCR (XP cycler, BIOER, Hangzhou, China) with the same primers that were used for cloning.…”

Section: Pcr Analysis Of Transgenic Plantsmentioning

confidence: 99%

A Gene Encoding Scots Pine Antimicrobial Protein Sp-AMP2 (PR-19) Confers Increased Tolerance against Botrytis cinerea in Transgenic Tobacco

Jaber

Kovalchuk

Raffaello

et al. 2017

Forests

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Both the establishment of sustainable forestry practices and the improvement of commercially grown trees require better understanding of mechanisms used by forest trees to combat microbial pathogens. We investigated the contribution of a gene encoding Scots pine (Pinus sylvestris L.) antimicrobial protein Sp-AMP2 (PR-19) to the host defenses to evaluate the potential of Sp-AMP genes as molecular markers for resistance breeding. We developed transgenic tobacco plants expressing the Sp-AMP2 gene. Transgenic plants showed a reduction in the size of lesions caused by the necrotrophic pathogen Botrytis cinerea. In order to investigate Sp-AMP2 gene expression level, four transgenic lines were tested in comparison to control and non-transgenic plants. No Sp-AMP2 transcripts were observed in any of the control and non-transgenic plants tested. The transcript of Sp-AMP2 was abundantly present in all transgenic lines. Sp-AMP2 was induced highly in response to the B. cinerea infection at 3 d.p.i. This study provides an insight into the role of Sp-AMP2 and its functional and ecological significance in the regulation of plant-pathogen interactions.

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“…Genomic DNA was extracted from the control and experimental lines according to the standard technique (СТАВ) [18].…”

Section: Southern Blotting Analysismentioning

confidence: 99%

Genetically Transforming Russian Potato Cultivars for Resistance to Colorado Beetle

Kamionskaya¹,

Кузнецов²,

Ismailov³

2012

Clon Transgen

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Using Agrobacterium-mediated transformation procedure, a total of 204 primary transformants of the Elizaveta, Lugovskoi and Nevski potato varieties carrying the cry IIIa gene were obtained. A number of biomolecular tests, including Southern hybridization, insert copy number screening, immunofluorescence analysis for assessment of the target gene expression levels, and PCR control for testing of the insert integrity, was carried out for these transformants, as well as the biosafety field trials for the transgenic lines selected during the biomolecular analysis. As a result, 3 transgenic lines (E2, N1, and L5) carrying one insert copy per genome with target gene expression level over 10 ppm were selected.

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Preparation of Genomic DNA from Plant Tissue

Cited by 150 publications

References 6 publications

A novel and efficient protocol for the isolation of genomic DNA from<br>mulberry (Morus L.)

A novel and efficient protocol for the isolation of genomic DNA from<br>mulberry (Morus L.)

A Gene Encoding Scots Pine Antimicrobial Protein Sp-AMP2 (PR-19) Confers Increased Tolerance against Botrytis cinerea in Transgenic Tobacco

Genetically Transforming Russian Potato Cultivars for Resistance to Colorado Beetle

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