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Quality DNA Isolation Using Different Methods of Sugarcane (saccharum Officinarum L.)

M. A. Hossain
Bangladesh Sugarcane Research Institute, lshurdi, Pabna, Bangladesh.

M. M. Shaik
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia, Bangladesh.

R. M. S. Shahnawaz
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia, Bangladesh.

N. Islam
Bangladesh Sugarcane Research Institute, lshurdi, Pabna, Bangladesh.

M. R. Islam
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia, Bangladesh.

M.A. S. Miah
Bangladesh Sugarcane Research Institute, lshurdi, Pabna, Bangladesh.

Abstract/ Executive Summary:

Three methods were used for DNA isolation from fresh leaf tissues (laminar tissue and meristem cylinder) collected from field grown 8 month old plants. The samples were grind without liquid nitrogen. The quality and quantity of DNA were determined by visu al estimation of electrophoreses agarose gel and by spectrophotometer. Method I included use of Ultra-Turax for sample grinding, refrigerated centrifuge for separation of different com ponents of samples and large amount chemicals were used in different steps compared to methods II and Ill. In both Method II and Method Ill grinding was done using mortar pestle in polypropylene bag at room temperature. Quality and quantity of DNA were determined by electrophoreses gel observation staining with Ethidium bromide, and spectrophotometer. Different DNA amounts were recorded in different isolation methods. The highest amount of DNA was obtained from the Method Ill (4100-4600 ngml-1) followed by Method II (1400-1950 ngml-1) and Method-! (1250-1650 ngml-1). Quality DNA isolation was possible without use of liquid nitrogen, tissue homoginizer and using a simple micro-centrifuge.

Keywords: Sugarcane, Quality DNA, DNA isolation

Location: Bangladesh Sugarcane Research Institute, lshurdi, Pabna, Bangladesh.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Sugarcane

Objectives:

To isolate polysaccha rides rapidly and polyphenol-free pure genomic DNA from sugarcane using low cost equipment as well as avoiding liquid nitrogen.

Methodology:

In this experiment two types of samples such as meristem cylinder and Top Visible Dulap (TVD) leaves of sugarcane were used. Top of the field grown 8 month old sugarcane plants was cut and placed vertically in a bucked with water to preserve alive and fresh. In laboratory, for meristem cylinder outer leaf sheaths were removed leaving inner spindle. Then the spindle base was cut away with sterile scissors and required amount was taken using a fine balance. TVD leaf was taken and mid rib was discarded. Then middle portion of leaf blade was cut into pieces of about 1cm x 1cm size. Then required amount was weighed with a fine balance.

Source of Information: Bangladesh Journal of Sugarcane, Vol: 28, pp: 65-69, July, 2006, ISSN 1991-8925

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The findings of the present study indicate the highest amount of DNA was obtained from the Method Ill (4100-4600 ngml-1) followed by Method II (1400-1950 ngml-1) and Method-I (1250-1650 ngml-1). Quality DNA isolation was possible without use of liquid nitrogen, tissue homoginizer and using a simple micro-centrifuge.

Knowledge Management: Journal