Abstract

BERMUDEZ-GUZMAN, Manuel de Jesús et al. Optimizing a protocol for DNA isolation of leaf Saccharum officinarum. Rev. Mex. Cienc. Agríc [online]. 2016, vol.7, n.4, pp.897-910. ISSN 2007-0934.

The DNA extraction free polysaccharides, polyphenols, plant protein and RNA is the initial step to several studies in molecular biology. There are various methods of extracting specific DNA of sugar cane (Saccharum officinarum), however, are time consuming and expensive equipment and reagents used. The aim of this study was to optimize a fast, efficient and inexpensive protocol for extracting DNA from leaves of S. officinarum. The variables evaluated were yield, purity, integrity and functionality of DNA purified from leaves of sugarcane 6 months old. Techniques spectrophotometry, agarose gel electrophoresis and molecular markers to evaluate the above variables were used. The performance variables (mg/g) and purity (A260:280 and A260:230) the extracted DNA showed highly significant differences (p˃ 0.05) protocol I. The best results were obtained with the protocol interaction I +N2, DNA yield 1.17 mg/g and 1.95 and 1.91 purities (A260:280 and A260:230). Functionality DNA by PCR amplification with the RAPD markers and TRAP bands sharp profiles generated good quality and easy to interpret. The use of the protocol I+N2 for extracting DNA from S. officinarum to obtain high yields of DNA with optimal quality and purity for applications of molecular markers is recommended.

Keywords : Saccharum officinarum; CTAB; liquid nitrogen; molecular markers.

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