PJB-2019-685
Standardization of different protocols for genomic DNA isolation from Phoenix dactylifera L.
Shamsa Kanwal, Shakra Jamil, Rahil Shahzad, Sajid-Ur-Rahman and Muhammad Zaffar Iqbal
Abstract
DNA fingerprinting, gene sequencing, genetic diversity studies and other valuable applications in molecular biology require highly purified DNA isolation for further use. DNA isolation is quite difficult from the hard and fibrous leaves of date palm. Tissues also contain high concentration of polyphenolics and polysaccharides linked with genomic DNA that may interfere with PCR amplification. This study was aimed at selection of the most appropriate method for DNA isolation from fibrous tissues of date palm and finding the alternative ways of grinding leaf samples to evade the problems linked with availability and usage of liquid nitrogen in date palm growing areas. DNA extraction from leaves of five date palm varieties was accomplished using different protocols including modified DNAzol method, cetyltrimethylammonium bromide (CTAB) method: (a) using liquid nitrogen for sample grinding and (b) grinding of leaf samples using sterile sand. Three different lysis buffers with different constituents were used. Study was carried out to observe the sodium chloride (NaCl) effects on produce and pureness of DNA individually and in combination with polyvinylpyrrolidone, (PVP) and lithium chloride (LiCl). It was concluded that DNA extracted using DNAzol was best in purity and yield. It was observed that crushing the leaf samples of date palm using sterile sand along with addition of lysis buffer containing NaCl (1.4 M) without addition of liquid nitrogen, or PVP and LiCl, gave adequate DNA yield and satisfactory purity.
To Cite this article:
Kanwal, S., S. Jamil, R. Shahzad, S.U. Rahman and Muhammad Zaffar Iqbal - Standardization of different protocols for genomic DNA isolation from Phoenix dactylifera L. Pak. J. Bot., 53(5): DOI: http://dx.doi.org/10.30848/PJB2021-5(6)
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