Dr. Shamsa kanwal
AgriculturalBiotechnology Research Institute, Ayub AgriculturalResearch Institute, Faisalabad Pakistan
Title: Standardization of different protocols for isolation of high quality genomin dna from phoenix dactylifera l.) And effect of sand grinding and different constitution of lysis buffer
Biography:
Abstract:
DNA fingerprinting, sequencing of genes, use of molecular markers for genetic diversity analysis and other applications in molecular biology and biotechnology require highly purified DNA isolation for further use. Leaves of date palm are fibrous and rigid therefore genomic DNA isolation from these leaves is quite difficult. 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 best suitable method for DNA isolation from fibrous tissues of date palm and finding the alternative ways of grinding the 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 completed using different protocols including modified DNAzol method, cetyltrimethylammonium bromide (CTAB) method: (a) grinding with liquid nitrogen and (b) grinding of leaf samples using sterile sand and three different lysis buffers with different constituents. Individual and combined effects of sodium chloride (NaCl), polyvinylpyrrolidone (PVP) and lithium chloride (LiCl) for a DNA yield of adequately high purity and PCR amplification were assessed in this study. It was concluded that DNA extracted using DNAzol was best in purity and yield and it was observed that grinding of date palm leaves with sterile sand and addition of NaCl (1.4 M) in the lysis buffer without the expensive use of liquid nitrogen, PVP and LiCl, provides a DNA yield of sufficient purity, appropriate for PCR amplification.
Shamsa kanwal
AgriculturalBiotechnology Research Institute, Ayub AgriculturalResearch Institute, Faisalabad Pakistan
Title: Standardization of different protocols for isolation of high quality genomin dna from phoenix dactylifera l.) And effect of sand grinding and different constitution of lysis buffer
Biography:
Abstract:
DNA fingerprinting, sequencing of genes, use of molecular markers for genetic diversity analysis and other applications in molecular biology and biotechnology require highly purified DNA isolation for further use. Leaves of date palm are fibrous and rigid therefore genomic DNA isolation from these leaves is quite difficult. 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 best suitable method for DNA isolation from fibrous tissues of date palm and finding the alternative ways of grinding the 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 completed using different protocols including modified DNAzol method, cetyltrimethylammonium bromide (CTAB) method: (a) grinding with liquid nitrogen and (b) grinding of leaf samples using sterile sand and three different lysis buffers with different constituents. Individual and combined effects of sodium chloride (NaCl), polyvinylpyrrolidone (PVP) and lithium chloride (LiCl) for a DNA yield of adequately high purity and PCR amplification were assessed in this study. It was concluded that DNA extracted using DNAzol was best in purity and yield and it was observed that grinding of date palm leaves with sterile sand and addition of NaCl (1.4 M) in the lysis buffer without the expensive use of liquid nitrogen, PVP and LiCl, provides a DNA yield of sufficient purity, appropriate for PCR amplification.
Shamsa kanwal
AgriculturalBiotechnology Research Institute, Pakistan
Title: Standardization of different protocols for isolation of high quality genomin dna from phoenix dactylifera l.) And effect of sand grinding and different constitution of lysis buffer
Biography:
Abstract:
DNA fingerprinting, sequencing of genes, use of molecular markers for genetic diversity analysis and other applications in molecular biology and biotechnology require highly purified DNA isolation for further use. Leaves of date palm are fibrous and rigid therefore genomic DNA isolation from these leaves is quite difficult. 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 best suitable method for DNA isolation from fibrous tissues of date palm and finding the alternative ways of grinding the 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 completed using different protocols including modified DNAzol method, cetyltrimethylammonium bromide (CTAB) method: (a) grinding with liquid nitrogen and (b) grinding of leaf samples using sterile sand and three different lysis buffers with different constituents. Individual and combined effects of sodium chloride (NaCl), polyvinylpyrrolidone (PVP) and lithium chloride (LiCl) for a DNA yield of adequately high purity and PCR amplification were assessed in this study. It was concluded that DNA extracted using DNAzol was best in purity and yield and it was observed that grinding of date palm leaves with sterile sand and addition of NaCl (1.4 M) in the lysis buffer without the expensive use of liquid nitrogen, PVP and LiCl, provides a DNA yield of sufficient purity, appropriate for PCR amplification.