از گياه انار DNA مقايسه شش روش استخراج

COMPARISON OF SIX GENOMIC DNA EXTRACTION METHODS FROM POMEGRANATE (PUNICA GRANATUM L.)

Genomic DNA extraction with a high quantity and quality is one of the most important primary needs in genomics laboratories. Extraction of DNA from woody plants, such as fruit trees, besides the presence of polysaccharides, is faced with the problem of phenolic compounds, which negatively affects the DNA quality. Therefore, use of methods for DNA extraction to be able to reduce these compounds to the lowest level is highly preferred. In order to select the best genolnic DNA extraction method from pomegranate plant tissues, which is distinct in phenolic compounds, six DNA extraction methods were compared. The methods were: 1- Vroh Bi et at (1996), 2- Dellaporta et al. (1983). 3-Ziegenhagen et al. (1993). 4- Doyle and Doyle (1990). 5- Lodhi et al. (1994), 6- Murry and Thompson (1980). Young, mature and old leaves and also peel and aril of fruits were used as plant materials for DNA extraction. Quantity and quality of extracted genomic DNAs were compared using the spectrophotometric, agarose gel electrophoresis, digestion by restriction enzymes and polymerase chain reaction (PCR) methods. Ultimately, considering the quantity and quality of DNA. the method of Vroh Bi et al. for young, mature and old leaves and also fruit parts and the method of Murry and Thompson for young leaves were chosen. Highest amount of DNA (64.7 µg) from 1 g young leaf tissues with good quality was obtained using Vroh Bi et al. method, which was not significantly different from the amount of extracted DNA (62.4 µg) from 1 g mature leaves. According to the results, the method of Vroh Bi et al. is recommended for DNA extraction from different tissues of plants having high level of polysaccharides and phenolic compounds.