Evaluation of Efficiency of Different DNA Extraction Protocols and Molecular Identification of Commercial Soybean (Glycine max. , L.) Growing in Turkey

Abstract

Four DNA extraction methods were used to obtain DNA from herbarium specimens of soybean and fresh soybean seeds, and germinated soybean seeds in this study. The quality of DNA obtained was estimated by using a spectrophotometer to measure the A260/280 absorbance ratio. Four different DNA extraction methods were compared for the isolation of DNA from the soybean homogenates, namely the CTAB extraction method, Plant Genomic DNA Purification Kit method,EZ1 Nucleic acid isolation method, and DNA extraction with phenol purification and liquid nitrogen. The main goal of study is to evaluate various methods of DNA isolation in terms of DNA yield and amplification quality. To preserve DNA well, it is necessary to dry plants as fast as possible. Extraction results depend on how the plant material is prepared, and the type of chemicals or DNA isolation protocols used. Obtaining high quality DNA depends on the isolation technique used. Several methods that are useful for dry plant tissue from herbarium specimens have been described [9, 16, 19]. Four extraction protocols were compared using fresh material and dry herbarium specimens and seeds. Plant Genomic DNA Purification Kit method; the concentration of DNA in the second elution extract was very low, thus repeated elution was not applied. DNA extraction is done with phenol purification and liquid nitrogen. This technique is not quite suitable for fresh plant leaves. It is usually used for herbarium samples of at least 0.2 g CTAB extraction method. This protocol is quite suitable for DNA isolation from the fresh plant material or germinated seed plantlets. The total DNA was isolated from approximately 0.5-1 g of fresh leaves (conserved in CTAB) of the same collection of sample. EZ1 Nucleic acid isolation tehnique is quite useful for high yield and quality of DNA isolation from dried soybean seeds. In this methods, no further purification was needed for molecular analysis

Keywords

• Genomic DNA extraction, soybean (Glycine max. L.)