An efficient method to extract DNA from refined rapeseed oil

Genetically modified oilseeds were used as processing raw material for edible oils. The protection of consumer rights to information as well as the genetically modified orgamisms （GMO） labe-ling polily required analytical methods to determine whether the oils contained genetically modified ingre-dient or not. Polymerase chain reaction （PCR） - based method was used commonly to determine the presence of GMOs. Adulteration attracted peoples concern also. Thus it was crucial that enough DNA ex-tracts can be obtained successfully from oil samples. For the purpose, three DNA extraction methods （modified emulsification method, the kits Wizard Magnetic DNA purification system for food and Nucleospin Food） ,were applied to 3 different grades of rapeseed oil samples. Those methods were compared by the amplification of Brassica napus reference gene CruA using real - time PCR. The results demonstra-ted that both the modified emulsification and the Nucleospin methods were able to extract enough DNA from refined oils. The modified emulsification method was superior to the kit of Nucleospin food due to smaller volume of required sample.

A Method Suitable for Extracting Genomic DNA from Animal and Plant——Modified CTAB Method

[Objective] The study aimed to introduce a rapid and effective method that is suitable for extracting genomic DNA from animal and plant. [ Method ] The genomic DNAs were extracted from tender leaves of 24 peanut cuhivars and from the liver, lung and kidney of white mouse through the specifically modified CTAB method. The DNAs were run on agarose gel, next detected by DNA/Protein analyzer. Finally PCR amplification was conducted to detect the quality of DNAs extracted using the modified CTAB method. [ Result] The clear and orderly bands were observed in gel detection, and the values of OD200/OD200 for DNAs extracted via modified CTAB method were between 1.77 - 1.83. The DNAs performed well in PCR amplification. [ Conclusion] The DNAs extracted by modified CTAB method could satisfy the requirement of PCR amplification.

Modified CTAB Method for Extracting Genomic DNA from Wheat Leaf

ObjectiveThe aim was to seek for a rapid DNA minipreparation method from wheat leaf. MethodThe total DNA of wheat leaf was extracted using CTAB, SDS and boiling water, separately, with some modifications. Integrity and purity of nucleic acids were detected through agarose gel electrophoresis, ultraviolet absorption and PCR. ResultThe DNA extracted by the modified CTAB method had high quality and purity, and was not degraded. Two hundreds of DNA samples could be extracted each workday by per capita using this method; and the PCR detection of wheat transgenic plants showed that amplified bands of target gene were clear, without false-positive, and the test results were satisfactory. The DNA purity and concentration extracted by modified SDS method were not as good as that extracted by modified CTAB method, but it also met the DNA requirements of major molecular research. The DNA quantity extracted by modified boiling method was small and there were a lot of impurities in it, PCR detection of this DNA showed no amplified band. ConclusionModified CTAB method is a simple and rapid method for DNA minipreparation from wheat leaf, and was suitable for PCR amplification and other molecular biology researches.

A Simple and Quick Method of Extracting Genomic DNA from Wheat Leaves

[ Objective] The aim was to seek a simple and quick method of extracting genomic DNA from wheat leaves. [ Method] Taking tender leaves of wheat as test materials, total DNA of transgenic wheat was extracted by using modified CTAB method. The extracted DNA was detected by 0.8% agarose gel electrophoresis. [ Result] DNA purity of extracted genome DNA from wheat was high and no degradation phenomenon using modified CTAB method, and was suitable for carrying out normal PCR amplification. [ Conclusion] This study provides a simple and quick method for extracting DNA from wheat with a spot of material.

An Improved Method of Extracting Artemisia abrotanum Genomic DNA

[ Objective ] The objective of this study is to explore a rapid and efficient method of extracting genomic DNA from Artemisia abrotanum. [ Method] Three methods of Cutting Method, Liquid Nitrogen Method and Quartz Sand Method based on SDS method were employed to extract Artemisia abrotanum genomlc DNA from tender leaf at seedling stage, tender spike and old leaf at heading stage. The obtained DNAs were detected by absorbance detection, agarose gel and PCR amplification. [ Result ] Cutting Method performed better than the other two methods compared in purity, extracting cycle and cost, accordingly more suitable for PCR amplification. The results also show that young spike is the best material for extracting genomic DNA from Artemisia Annua.

A Rapid DNA Extraction Method for PCR Detection of Arabidopsis thaliana

[Objective] The aim was to introduce a rapid DNA extraction method for PCR detection of Arabidopsis thaliana.[Method] Through the improvement of conventional DNA extraction method,a rapid Arabidopsis thaliana DNA extraction method was obtained.With randomly selected Arabidopsis thaliana transgenic strains and mutants as samples,the method was verified.[Result] After electrophoresis,UV absorption detection,it was found that DNA samples are complete and less pollution,and the result of PCR amplification objective fragment was good which proved DNA is suitable as a template for PCR reaction.After PCR detection,positive plants gene amplified bands were clear,without false-positive,and the test results were satisfactory.[Conclusion] The method is suitable for rapid extraction of Arabidopsis thaliana DNA and PCR detection.

Isolation of Chlorella vulgaris and Its DNA Extraction Methods

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.

Comparison of Genomic DNA Extraction Methods for Chenopodium quinoa Willd

To rapidly obtain high-quality genomic DNA from Chenopodium quinoa Willd, the genomic DAN in different tissues （leaves, stems and roots） of Chenopodi- um quinoa Willd was extracted by modified CTAB method, SDS method and high- salt Iow-pH method, respectively. The quality and yield of extracted DNA was deter- mined using agarose gel electrophoresis and UV spectrophotometry. At the same time, the PCR-SSR and SSCP molecular detection was also performed. The results showed that the gel test strips, without obvious decomposition, of all the extraction methods were relatively obvious; the genomic DNA yield extracted by modified CTAB method was highest, followed by that by SDS method, and the genomic DNA extracted by high-salt Iow-pH method was lowest： the genomic DNA yields extracted by different methods from Chenopodium quinoa Wiltd leaves were all high- er than those from roots and stems; the quality of Chenopodium quinoa Willd ge- nomic DNA extracted by modified CTAB method and high-salt Iow-pH method was better, and polyphenols, polysaccharides and other impurities were removed more completely. The PCR-SSR and SSCP detection results showed that the genomic DNA extracted by different methods from different tissues of Chenopodium quinoa Willd all could be better amplified, and high-quality strips could be obtained. So the Chenopodium quinoa Willd genomic DNA extracted by the three methods all can be used for subsequent molecular biology research.

Comparison on Four Extraction Methods of Genomic DNA from Clematis fasciculiflora Franch

[Objective] This study aimed at comparing the four extraction methods of genomic DNA from Clematis fasciculiflora Franch and determining the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch.[Method] Leavies of Clematis fasciculiflora Franch were used as materials for comparing the purity and concentration of extracted DNA and extracting time among the four extraction methods of genomic DNA including improved CTAB method Ⅰ,improved CTAB method Ⅱ,improved CTAB method Ⅲ and improved SDS method.[Result] The four extraction methods could all be successfully used for extracting the genomic DNA from Clematis fasciculiflora Franch.The purity of genomic DNA was the highest using improved CTAB method Ⅰ,with the longest extracting time;while the concentration of genomic DNA was the maximum using the improved SDS method,with the shortest extracting time and relatively low purity;the extracting time of improved CTAB method Ⅲ was the shortest.[Conclusion] This study had established the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch and supported for the further research using molecular biological methods.

Comparison on Methods.for Extracting DNA from Pteridophyta

[ Objective] The aim was to study method for extracting DNA from pteridophyta and provide basis for further study on genetic diversity and taxonomy. [ Method] By changing the dosage of reagent and operating method, CTAB method for extracting DNA was improved. [ Result] The results showed that the improved CTAB method could extract high-quality DNA from pteridophyta. [ Conclusion] The study improved method for extracting DNA from pteridophyta.

Genomic DNA Isolation by Phenol/Chloroform Extracting Method from Sheep Blood Clot

[ Objective] The aim was to establish the method of extracting genomic DNA from sheep blood clot on the basis of the improvement of method for extracting genomic DNA from tissues. [Method]The genomic DNA with complete primary structure and high purity was obtained from the sheep blood clot after the steps of cutting the sheep blood clot with ophthalmic scissors, cell lysis with tissue DNA extracts and digested by proteinase K, extracting with phenol/chloroform and precipitating with ethanol were performed. [ Result] The concentration of the extracted DNA was 159.90 ±0.70 ng/μl and the ratio of the A260/A280 was 1.80 ＋0.01. The sheep microsatellite locus of BM203 was amplified by using the extracted DNA from the sheep blood clot as template of PCR, and the PCR result was perfect. [Conclusion]This method is simple and feasible, the quantity and quality of the extracted DNA can satisfy the demands for the subsequent researches. It is worth to extending and using for reference.

A Rapid Metagenomic DNA Extraction from Sediments: Potassium Dichromate SDS Method

[Objective] The objective of this study was to report an improved method for rapid DNA extraction from black-order sediments, without any purification step. [Methods] Sediments in eutrophic lake are complex ecosystems and soil microbes involved in anthropogenic nutrient cycling are very important. DNA-based molecular methods offer new tools for characterization of these mixed communities of mi- croorganisms. However, it is very difficult to remove humic substances, heavy met- als that co-existed with genome DNA representing the microbial community directly from these complex systems and can interfere with subsequent genetic analysis. The potassium dichromate solution was firstly used to remove humic substances. [Results] The steps of removing humic substances and DNA extraction were per- formed simultaneously that improved the speed of extraction to approximately 1 hour and the nucleic acids that were obtained with this method did not need to be washed with 70% ethanol and dissolved directly in sterile water for total bacterial 16S rDNA, nosZ gene of denitrifying bacteria, pmoA of methanotrophs, nifH of nitro- gen-fixing bacteria, amoA of ammonia-oxidizing bacteria and ammonia-oxidizing ar- chaea molecular ecology analyses. [Conclusion] This method could provide a plat- form for preparing a fast sediments DNA extraction.

Comparison on Several Methods for Extracting DNA from Raccoon Dog Hair Follicle

[Objective] This study aimed to investigate a reliable method for DNA ex- traction from Wusuli raccoon dog＇s hair. [Method] Several DNA extraction methods were used to extract DNA from Wusuli raccoon dog hair, including Chelex-100 method, PCR buffer method, organic phenol-chloroform method and centrifugal col- umn type kit method. The extracted DNA was analyzed by using PCR amplification and electrophoresis to compare these four DNA extraction methods. [Result] Accord- ing to the results of spectrophotometer detection and gel electrophoresis, nucleic acid extracted by Chetex-100 method had proteins and other impurities; nucleic acid ex- tracted by PCR buffer method was low in concentration; however, DNA extracted by organic phenol-chloroform method and centrifugal column type kit was high in con- centration with no impurity band. [Conclusion] This study had laid the strong founda- tion of scientific theory to further explore the efficient and simple method for extracting DNA from Wusuli raccoon dog hair follicle.

Rapid DNA Extraction Method from Amorphophallas.konjac and ISSR-PCR Amplification

[Objective]The research aimed to study the total DNA extraction methods of Amorphophallas.konjac.[Method]To investigate the optimum DNA extraction method,CTAB,SDS and new modified methods for isolating genomic DNA from Amorphophallas.konjac leaf were studied and the isolated DNAs were examined by ultraviolet absorption test,agarose gel electrophoresis and ISSR-PCR test for comparative analysis.[Result]The results showed that the new modified method was suitable for DNA isolation from Amorphophallas.konjac plants with high purity and yield;quality of DNA could meet ISSR-PCR requirement even extracted once by the new modified method.[Conclusion]The study provided basis for extracting high quality DNA sample,and offered the molecular genetics reference for exploring the genetic diversity of Amorphophallas.konjac.

DNA Extraction and Molecular Identification of Green Tea

[Objective] The aim of this study was to provide reference for the quality identification of green tea.[Method] Green Tea was used as materials,and its total DNA was extracted through improved CTAB method.And the obtained DNA was used to carry out identification on 10 varieties of green tea through ISSR molecular markers.[Result] The high quality DNA from green tea could be obtained with new method,the DNA yield ranged from 101-498 μg/g tea sample for various green tea samples,and the average yield was 249 μg/g tea sample.The ISSR detection result showed that ISSR markers could effectively differentiate different varieties of green tea.[Conclusion] The result had provided reference for the further study on molecular identification of green tea.

A New Rapid and Batch-oriented Crushing Method for DNA Extraction from Maize Leaves

The crushing of leaves is the most time and effort-consuming part in DNA extraction which is a fundamental step in the study of molecular biology. In this study, a new rapid and batch-oriented crushing method for DNA extraction from maize leaves was developed. In addition, the practicability of the developed method in molecular marker-assisted breeding was verified using SSR molecular maker technology so as to provide a rapid, batch-oriented, low-cost and non-toxic leafcrushing method for a large number of molecular marker tests, improving test efficiency.

An Improved Method for DNA Extraction from the Faeces of Red Deer

[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the characteristics of red deer faeces. [Result] This improved method extracted high-quality fecal DNA from Tianshan red deer and amplified the mitochondrial cytochrome b gene. With the muscle and fur DNA of red deer as the control, the sequencing results further con- firmed the reliability of the method. [Conclusion] The method requires no proteinase K in the process of extraction, and the extracted DNA can be used for PCR ampli- fication directly without the purification of DNA purification kit, thus, it is cost-saving.

Methodological Studies on Genomic DNA Extraction and Purification from Plant Drug Materials

This paper reports the rationale and methods of DNA extraction and purification from nine species of Compositae and four commercial drugs of corresponding plant Elephantopus scaber. The comparison of three methods: CsCl gradient, CTAB/CsCl gradient and CTAB miniprep extraction by yield, purity and factors affecting PCR was carried out. In conclusion, CTAB miniprep method provides a rapid, effective, economic approach for isolating genomic DNA for Chinese drug identification by genomic fingerprints.

A Mass and Rapid Method for DNA Extraction of Beauveria bassiana

[Objective] The aim was to optimize the mass and rapid method for DNA extraction of Beauvena bassiana. [Method] Boiling water DNA extraction method was improved, DNA extraction liquid was heated by PCR instrument and the extraction process was finished rapidly. [ Resuit] The quality of DNA obtained through mass and rapid extraction of fungal genomic DNA could meet the requirement of RAPD amplification analysis. The clear bands were amplified from 22 tested strains, the number of clear bands were different in the range of 2 -6 and the size of band were mainly concentrated in 450 -800 bp. The DNA extracted by this method also could completely meet the requirement of SCAR amplification. The amplified specific DNA bands used to mark the strain F263 were very clear. [Conclusion] This research provided relatively perfect method for mass and rapid extraction of fungal clenomic DNA.

“DNA粗提取和鉴定”实验设计与改进

对DNA粗提取与鉴定实验进行优化,探究植物组织的多种破碎方法对DNA粗提取的影响,并用氯仿等有机物对DNA粗提取物进一步纯化,分析DNA未纯化与纯化后的纯度差异。结果显示,相较于破壁机破碎、加液氮研钵研磨以及玻璃匀浆器研磨,不添加液氮用研钵研磨效果更好。DNA粗提液经纯化后纯度显著提高,蛋白质及盐类等污染明显减少。