A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Bo...A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Boom et al. (1990). By modifying the protocol, plant genome DNA could be extracted easily from dormant buds, mature leaves, and other parts of plant. Our results showed that the purified DNA was of high purity and could be analyzed by PCR. Furthermore, this CTAB-silica method took much less time for a successful DNA purification process compared to the traditional methods (CTAB and SDS). By our method, the suitable DNA can be extracted and purified from over 10 plant samples by one person in an hour.展开更多
The sorption behavior of acid dyes onto cetyltrimethylammonium bromide (CTAB)-modified silica as a function of pH in the aqueous medium was studied. Single-and multi-solute sorption equilibria of orange Ⅱ（OR), pheno...The sorption behavior of acid dyes onto cetyltrimethylammonium bromide (CTAB)-modified silica as a function of pH in the aqueous medium was studied. Single-and multi-solute sorption equilibria of orange Ⅱ（OR), phenol red (PR) and Eriochrome Black T (EBT) were studied at pH 3, unbuffered water pH and pH 11. Sorption behavior of EBT could not be conducted at pH3 due to its aggregation in acidic medium. All the reaction conditions, experimental protocols and techniques remained the same throughout the sorption process. Sorption isotherms for single-solute system were fitted by the Langmuir model, while Langmuir competitive model (LCM) and the ideal adsorbed solution theory (IAST) coupled with Langmuir model (IAST/Langmuir) were used for the prediction of multisolute competitive sorption. Sorption affinities influenced by the factors like physical interactive forces between the molecules of CTA on silica and sorbate, structural limitations of the dyes based on their geometrical arrangement were investigated. Sorption affinity of OR was found to be higher than that of EBT and PR at all the pH values investigated. Magnitude of the sorption capacities was observed to be higher in acidic medium but lower in alkaline medium. Trends of the sorption affinities in multisolute system were simlar to those in single-solute system but magnitude of the sorption capacities was significantly reduced due to the prevailing competition among the sorbates.展开更多
Isolation of high quality DNA from multiple samples can be both time-consuming and expensive. We have developed a combined protocol to reduce the time component of the hexadecyltrimethylammonium bromide （CTAB） extra...Isolation of high quality DNA from multiple samples can be both time-consuming and expensive. We have developed a combined protocol to reduce the time component of the hexadecyltrimethylammonium bromide （CTAB） extraction method and reduced costs by regenerating the silica columns used to purify genomic DNA. We present data that shows, by in- creasing the temperature used during the CTAB method, the time required to extract crude genomic DNA can be reduced. We show that silica columns can be regenerated using HCI and still maintain their DNA-binding capacity. Furthermore, we show both spectrophotometrically, and by restriction enzyme cutting, that the quality of the eluted DNA is high. Critically, using both genomic DNA from pea and perennial ryegrass we demonstrate, using species-specific PCR primers, that there is no carry-over of DNA from repeated use of a single column. The main advantages of the method are high yield, high quality, cost effectiveness and time-saving. This method could satisfy demand when large numbers of plant genomic DNA samples are required, for example from targeting induced local lesions in genomes （TILLING） populations.展开更多
文摘A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Boom et al. (1990). By modifying the protocol, plant genome DNA could be extracted easily from dormant buds, mature leaves, and other parts of plant. Our results showed that the purified DNA was of high purity and could be analyzed by PCR. Furthermore, this CTAB-silica method took much less time for a successful DNA purification process compared to the traditional methods (CTAB and SDS). By our method, the suitable DNA can be extracted and purified from over 10 plant samples by one person in an hour.
文摘The sorption behavior of acid dyes onto cetyltrimethylammonium bromide (CTAB)-modified silica as a function of pH in the aqueous medium was studied. Single-and multi-solute sorption equilibria of orange Ⅱ（OR), phenol red (PR) and Eriochrome Black T (EBT) were studied at pH 3, unbuffered water pH and pH 11. Sorption behavior of EBT could not be conducted at pH3 due to its aggregation in acidic medium. All the reaction conditions, experimental protocols and techniques remained the same throughout the sorption process. Sorption isotherms for single-solute system were fitted by the Langmuir model, while Langmuir competitive model (LCM) and the ideal adsorbed solution theory (IAST) coupled with Langmuir model (IAST/Langmuir) were used for the prediction of multisolute competitive sorption. Sorption affinities influenced by the factors like physical interactive forces between the molecules of CTA on silica and sorbate, structural limitations of the dyes based on their geometrical arrangement were investigated. Sorption affinity of OR was found to be higher than that of EBT and PR at all the pH values investigated. Magnitude of the sorption capacities was observed to be higher in acidic medium but lower in alkaline medium. Trends of the sorption affinities in multisolute system were simlar to those in single-solute system but magnitude of the sorption capacities was significantly reduced due to the prevailing competition among the sorbates.
基金funding and the PhD Scholarship for Fu Zeyu from the New Zealand Foundation for Arable Researchfunding for Song Jiancheng from the National Natural Science Foundation of China (31371616)
文摘Isolation of high quality DNA from multiple samples can be both time-consuming and expensive. We have developed a combined protocol to reduce the time component of the hexadecyltrimethylammonium bromide （CTAB） extraction method and reduced costs by regenerating the silica columns used to purify genomic DNA. We present data that shows, by in- creasing the temperature used during the CTAB method, the time required to extract crude genomic DNA can be reduced. We show that silica columns can be regenerated using HCI and still maintain their DNA-binding capacity. Furthermore, we show both spectrophotometrically, and by restriction enzyme cutting, that the quality of the eluted DNA is high. Critically, using both genomic DNA from pea and perennial ryegrass we demonstrate, using species-specific PCR primers, that there is no carry-over of DNA from repeated use of a single column. The main advantages of the method are high yield, high quality, cost effectiveness and time-saving. This method could satisfy demand when large numbers of plant genomic DNA samples are required, for example from targeting induced local lesions in genomes （TILLING） populations.
