Comparison of DNA extraction methods from Geranium (Geraniaceae)

Abstract

The genus Geranium (Geraniaceae); with about 320 species throughout the temperate regions, is chemically characterised by the presence of tannins, flavonoids, anthocyanins and essential oils which interfere with the extraction of pure genomic DNA. It is necessary to optimise the extraction protocols to reduce the effects of the presence of these compounds to the lowest level. The present study compares the plant genomic DNA extraction Kit (DNP TM Kit), CTAB DNA extraction method by Murray and Thompson and Sahu et al., from the extracting DNA point of view Geranium species. The results showed significant differences in DNA contents between the three methods. Quantity and quality of extracted genomic DNAs were compared by employing the spectrophotometer, Nano-Drop, agarose gel electrophoresis, and polymerase chain reaction (PCR) methods and molecular marker such as (ITS and trnL-F) and ISSR. The method of Sahu et al., provided the best results (200 ng/µL) in terms of quantity and quality of DNA, therefore, this method was taken and optimised for DNA extraction. Our results proposed that this method could be effective for plants with same polysaccharides, proteins and polyphenols components. The advantage of this method is that it omits the use of liquid nitrogen and toxic phenols which are expensive. The success of this method in obtaining high-quality genomic DNA has been demonstrated in the Geranium species group and the reliability of this method has been discussed.

Genetic diversity and relationships among Hypericum L. species by ISSR Markers: A high value medicinal plant from Northern of Iran

“…These were dried by silica gel powder. CTAB activated char-coal protocol was used to extract genomic DNA (Esfandani- Bozchaloyi et al 2019). The quality of extracted DNA was examined by running on 0.8% agarose gel.…”

Evaluation of genetic diversity in geranium (Geraniaceae) using RAPD marker

“…Leaves were dried. DNA extraction was carried out according to the previous protocol (ESFANDANI-BOZCHALOYI et al, 2019). DNA quality was checked on an agarose gel to confirm the purity.…”

ISSR markers and population differentiations in Erodium ciconium (L.) L'hér ex Aiton

“…These were dried by silica gel powder. CTAB activated charcoal protocol was used to extract genomic DNA (Esfandani-Bozchaloyi et al, 2019). The quality of extracted DNA was examined by running it on 0.8% agarose gel.…”

“…Recent ISSR studies of natural populations have demonstrated the hypervariable nature of these markers and their potential use for population-level studies (Hulte´n and Fries, 1986). Limitations of the ISSR technique, as is the case for Random Amplification of Polymorphic DNA (RAPD; Esfandani-Bozchaloyi et al, 2019), are that the bands are scored as dominant markers and the genetic diversity estimates are based on diallelic characters. In the present study, ISSR markers were employed to analyze genetic diversity in 110 E. ciconium accessions belonging to 15 different populations for the first time in the Iran.…”