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High-quality plant DNA extraction for PCR: an easy approach

100%

Ahmed I., Islam M., Arshad W., Mannan A., Ahmad W., Mirza B.

Journal of Applied Genetics

2009

tom 50

nr 2

105-107

Polymerase chain reaction has found wide applications in modern research involving transformations and other genomic studies. For reproducible PCR results, however, the quantity and quality of template DNA is of considerable importance. A simple and efficient plant DNA extraction procedure for isolation of high-quality DNA from plant tissues is presented here. It requires maceration of plant tissue of about 1.0 cm² (e.g. of a leaf blade) in DNA extraction buffer (100 mM Tris-HCl, 100 mM EDTA, 250 mM NaCl) using 1.5-mL microfuge tubes, followed by cell lysis with 20% SDS, and DNA extraction with phenol: chloroform: iso-amyl alcohol (25:24:1 ). Hydrated ether is then used to remove polysaccharides and other contaminants from the DNA preparation. Average DNA yield is 20-30 µg cm⁻² for fresh tissues, and ratio of absorbance at 260 nm to absorbance at 280 nm is 1.5-1.8. The DNA is quite suitable for PCR using microsatellites, RAPD and specific markers for recombinant selection. Amplifications have been obtained for these markers by using template DNA extracted from fresh as well as frozen leaf tissues of various plants, including barley, oat, potato and tomato. DNA stored for more than 2 years has been successfully amplified with microsatellite markers, which shows suitability of this method after long-term storage of DNA. Besides, the ease of use and cost-effectiveness make the procedure attractive.

Gibberellic acid spray modulates fruiting, yield, quality, and shelf life of rambutan (Nephelium lappaceum L.)

100%

Uddin A. S. M. M., Gomasta J., Islam M. T., Islam M., Kayesh E., Karim M. R.

Journal of Horticultural Research

2024

tom 32

nr 1

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Boron facilitates rice growth, development, and related attributes under saline soil conditions

84%

Uddin R., Nuruzzaman M., Briste P. S., Islam M., Bhuiyan A. K., Joy I. H., Ahmed S., Khatun A.

Acta Agrobotanica

2021

tom 74

nr 1

Ograniczanie wyników

1 Acta Agrobotanica

1 Journal of Applied Genetics

1 Journal of Horticultural Research

3 Islam M.

1 Ahmad W.

1 Ahmed I.

1 Ahmed S.

1 Arshad W.

1 Bhuiyan A. K.

1 Briste P. S.

1 Gomasta J.

1 Islam M. T.

1 Joy I. H.

1 Karim M. R.

1 Kayesh E.

1 Khatun A.

1 Mannan A.

1 Mirza B.

1 Nuruzzaman M.

1 Uddin A. S. M. M.

1 Uddin R.

1 2024

1 2021

1 2009

Wyniki wyszukiwania

High-quality plant DNA extraction for PCR: an easy approach

Gibberellic acid spray modulates fruiting, yield, quality, and shelf life of rambutan (Nephelium lappaceum L.)

Boron facilitates rice growth, development, and related attributes under saline soil conditions