新型超声快速处理活检标本保存不同年限对DNA质量的影响

背景:新型超声组织处理技术越来越多地被用来进行分子生物学分析,研究新型超声处理不同存储年限组织DNA的质量,对进一步分子检测的标本质控具有重要意义。目的:探讨新型超声处理活检标本存储不同年限对DNA质量的影响,以期为分子检测探索最佳的标本存储时间。方法:收集40例乳腺穿刺小活检组织,采用超声技术制作石蜡标本,按照存储年限分为4组:<1年组、1-3年组、>3-5年组及>5年组,每组10例,对石蜡标本进行切片,每张切片厚3μm,切片10-15张,提取DNA后通过Nanophotometer N60超微量分光光度计和Qubit 4.0荧光计检测DNA的质量浓度,记录A_(260)/A_(280)比值判定DNA的纯度,利用全自动毛细管电泳核酸分析仪(Qsep 100)检测DNA片段完整性,以评估DNA片段的质量。结果与结论:4组样本A_(260)/A_(280)均值在1.8-2.0之间,达到纯度要求,无明显差异。4组样本的DNA质量浓度(Qubit浓度)均值分别为30.39,14.33,2.52,1.95 ng/μL;DNA的平均N/Q比值分别为6.48,14.18,24.56,29.86;DNA质量数均值分别为5.64,1.76,1.24,0.80;大片段占比均值分别为56.08%,17.72%,12.68%,7.90%。PCR检测内控基因Ct均值分别为15.32,17.09,18.39,21.24。与<1年组相比,其余3组DNA浓度显著降低,N/Q比值显著增加,DNA质量数和大片段占比均值显著降低,Ct值升高,差异有显著性意义(P<0.05)。实验结果表明,对于新型超声处理活检标本,应优先选择存储<1年的样本进行日常分子检测,储存3年内的样本可满足二代测序等检测要求,5年内样本仅可尝试进行PCR等检测,存储超过5年的样本不建议进行后续分子检测。

基于中医证候与精液质量相关参数构建精子DNA碎片预测模型与验证

背景:中医证候与精液质量相关参数相结合,共同预测精子DNA碎片指数(DNA fragmentation index,DFI)异常增高的发生并绘制列线图,能显著提高临床的实操性与应用效能,为临床全面评估精液质量,采取积极干预措施以改善临床结局及制定个体化医疗方案提供依据。目的:探讨基于中医证候与精液质量相关参数构建精子DNA碎片的预测模型与验证。方法:回顾性分析2019年7月至2021年7月在广西壮族自治区南溪山医院中医男科接受中医证候诊断及精子DNA碎片率检查的不育患者共420例,据《人类精液检查与处理实验室手册》(第6版),将其中137例精子DFI>30%患者纳入精子DFI异常增高组,将283例精子DFI≤30%作为对照组;首先采用单因素分析筛选精子DFI异常增高的影响因素,然后采用套索算法(LASSO)校正因子共线性问题并筛选出最佳匹配因子后,将其纳入多因素向前逐步Logistic回归找出其独立影响因素并绘制列线图,最后采用受试者工作曲线、校准曲线、临床决策曲线、临床影响曲线对该预测模型进行区分度与准确度及临床应用效能验证。结果与结论:①单因素分析结果显示,年龄、体质量指数、前向运动率、精子总活率、精子浓度、精子形态学、肾阳虚衰证、湿热下注证、肾精不足证为引发精子DFI异常增高的影响因子(P<0.05);②通过LASSO回归进一步筛选出的最佳匹配因素为年龄、体质量指数、精子总活率、精子浓度、精子形态学、肾阳虚衰证、湿热下注证、肾精不足证(P<0.05);③多因素向前逐步Logistic回归结果显示年龄、体质量指数、精子浓度、精子总活率、湿热下注证、肾阳虚衰证共6项为引发精子DFI异常增高的独立影响因素;④受试者工作曲线显示,模型组曲线下面积为0.760(0.713,0.806),验证组曲线下面积为0.745(0.714,0.776),说明该预测模型具有较好的区分度;⑤校准曲线平均绝对误差0.040,Hosmer-Lemeshow检验P>0.05,表明该模型预测发生精子DFI异常增高的概率与实际发生精子DFI异常增高的概率无显著统计学差异,证实该模型具有较好的准确度;⑥临床决策曲线与临床影响曲线显示,模型组与验证组分别在阈概率值为0.08-0.84与0.09-0.78时具有临床最大净获益,且在该阈概率范围内具有较好的临床应用效能;⑦结果表明,年龄、体质量指数、精子浓度、精子总活率、湿热下注证、肾阳虚衰证为引发精子DFI异常增高的独立影响因素,通过其构建的临床预测模型列线图具有较好的临床预测价值与临床应用效能,可为临床全面评估精液质量、预后与干预及个体化医疗服务提供依据。

基于活产建立体外受精-胚胎移植精子DNA碎片指数的参考阈值及子代短期安全性

背景:精子DNA碎片指数与受精、胚胎发育潜能、胚胎植入、流产及子代安全性等存在显著的相关性。然而,其临床参考值受多种因素的影响,导致临床意义极其有限,该研究以活产为结局,通过倾向评分匹配校正其他混杂因素后,构建精子DNA碎片指数与活产的最佳临床截断值,并对其进行内外部验证,具有较好的预测价值及临床应用效能。目的:探讨基于活产建立体外受精-胚胎移植精子DNA碎片指数的参考阈值及子代短期安全性。方法:选取2019年5月至2021年5月于常州市妇幼保健院接受体外受精-胚胎移植患者1921例,以倾向匹配容差0.02为标准,1∶1进行倾向评分匹配,结果活产组与非活产组各成功匹配540例,以此建立模型组;通过选取同时期广西壮族自治区南溪山医院接受体外受精-胚胎移植患者135例作为外部验证组;采用受试者工作曲线探求精子DNA碎片指数对活产的临床最佳截断值,分别采用限制性立方样条曲线、标准曲线、临床决策曲线、临床影响曲线及内外部验证等方法,对该截断值的准确性及临床应用效能进行评估。结果与结论:(1)非活产组精子DNA碎片指数显著高于活产组且与活产存在显著的负相关性(r=-0.444,P<0.001);(2)受试者工作曲线结果显示,DNA碎片指数对活产的最佳截断值为24.33%,曲线下面积为0.775(0.746,0.804),特异度为72.60%,敏感度为78.90%,准确度为75.70%;(3)限制性立方样条曲线拟合Logistic回归结果显示,当精子DNA碎片指数大于24.57%时,临床非活产的风险呈趋势性增涨;(4)Logistic回归概率分析结果显示,精子DNA碎片指数为活产的危险因素[OR(95%CI)=0.916(0.904,0.928),P<0.001],且当精子DNA碎片指数大于27.78%时,临床活产发生的概率将小于50%,随着精子DNA碎片指数每增高1个单位,活产的概率下降8.4%;(5)内外部对该临床截断值的验证均显示,该截点具有一定的临床预测价值及准确性;(6)临床决策曲线与临床影响曲线显示,以该临床截断值建立的预测模型在阈概率为0.22-0.73时具有临床最大净获益值,且在该阈概率范围内损失与获益的比值始终小于1,证实该预测模型具有较好的临床应用效能;(7)精子DNA碎片指数与子代短期安全性分析结果显示,精子DNA碎片指数与出生儿早产、体质量、畸形、性别差异无显著性;(8)结果表明,精子DNA碎片指数对体外受精-胚胎移植活产的最佳临床截断值为24.33%,以此建立的临床预测模型具有较好的区分度、准确度与临床应用效能,精子DNA碎片指数对子代短期安全性影响并不显著,但仍需大样本及长期的追踪评估。

FTA-环介导等温扩增技术直接提取变异链球菌DNA的效果评价

背景:变异链球菌是龋病的重要病原菌,及时检测变异链球菌水平对龋病的早发现、早治疗有重要意义。目的:建立并评价FTA-环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术直接提取变异链球菌DNA的应用效果。方法:①制备含有ATCC标准菌株变异链球菌的菌悬液,接种于脑心浸出液培养基,充分混匀后按10倍梯度稀释成7种浓度(4.2×10^(7),4.2×10^(6),4.2×10^(5),4.2×10^(4),4.2×10^(3),4.2×10^(2),4.2×10 CFU/mL),每个稀释级做2个平行对照,并增加无菌水作为空白对照;②分别采用FTA卡、常规煮沸法、试剂盒提取及裂解液提取4种方法直接提取菌株DNA,通过LAMP技术进行扩增,并进行特异性试验,比较4种提取方法的差异。结果与结论:①4种方法提取的DNA均满足LAMP扩增的要求;②特异性试验结果显示,只有变异链球菌才可特异扩增出靶基因;③裂解液提取法最低检测限为4.2×10^(3) CFU/mL,FTA卡提取法最低检测限为4.2×10^(4) CFU/mL,试剂盒提取法和常规煮沸法最低检测限分别为4.2×10^(6) CFU/mL和4.2×10^(7) CFU/mL;④4种提取方法其他方面的比较显示,试剂盒提取法的实验成本、步骤数和时间都是最高;其他3种方法步骤数一致,其中FTA卡所需仪器设备最少,常规煮沸法单次成本最低,裂解液提取法所需时间最少;FTA卡和裂解液提取法仅需少量菌即可提取成功,后者在时间方面优于FTA卡,但相较于FTA卡其单次成本高,所需设备多;⑤结果说明,该研究建立的FTA-LAMP技术具有操作简便、特异性强、灵敏度高、结果可视化等优势,有望为高效提取检测变异链球菌提供新途径。

Peripheral mitochondrial DNA as a neuroinflammatory biomarker for major depressive disorder

In the pathogenesis of major depressive disorder, chronic stress-related neuroinflammation hinders favorable prognosis and antidepressant response. Mitochondrial DNA may be an inflammatory trigger, after its release from stress-induced dysfunctional central nervous system mitochondria into peripheral circulation. This evidence supports the potential use of peripheral mitochondrial DNA as a neuroinflammatory biomarker for the diagnosis and treatment of major depressive disorder. Herein, we critically review the neuroinflammation theory in major depressive disorder, providing compelling evidence that mitochondrial DNA release acts as a critical biological substrate, and that it constitutes the neuroinflammatory disease pathway. After its release, mitochondrial DNA can be carried in the exosomes and transported to extracellular spaces in the central nervous system and peripheral circulation. Detectable exosomes render encaged mitochondrial DNA relatively stable. This mitochondrial DNA in peripheral circulation can thus be directly detected in clinical practice. These characteristics illustrate the potential for mitochondrial DNA to serve as an innovative clinical biomarker and molecular treatment target for major depressive disorder. This review also highlights the future potential value of clinical applications combining mitochondrial DNA with a panel of other biomarkers, to improve diagnostic precision in major depressive disorder.

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.

Comparative Study on Four Methods for Quick Extraction of Sorghum Genomic DNA

[Objective] This study was to find out a quick,simple,and low-cost method for the extraction of sorghum genomic DNA.[Method] Four plant genomic DNA extraction methods based on CTAB,including liquid nitrogen grinding method（method I）,buffer grinding method（method II）,drying grinding method（method III）and directly grinding method（method IV）,were used to extract the sorghum genomic DNA from leaves;further the quantity and quality of the yielded DNA were detected by gel electrophoresis,SSR-PCR and SRAP-PCR.[Result] These four methods performed no remarkable difference in DNA product.The method I and method II produced DNA with higher purity and better integrity,which,especially from method I,is effective for SRAP-PCR and SSR-PCR.While the DNA extracted via method III and method IV had less integrality and lower purity,and only effective in SSR-PCR.[Conclusion] Enough amount of sorghum genomic DNA to perform tens of PCR could be quickly extracted using all these four methods.The DNA obtained via method I and method II had a broader application spectrum（SRAP,RAPD,ISSR and SSR）than that via method III and method IV which is only proper for PCR targeting small DNA fragments（SSR）.

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.

The Distribution of Repetitive DNAs Along Chromosomes in Plants Revealed by Self-genomic in situ Hybridization

The distribution of repetitive DNAs along chromosomes is one of the crucial elements for understanding the organization and the evolution of plant genomes. Using a modified genomic in situ hybridization （GISH） procedure, fluorescence in situ hybridization （FISH） with genomic DNA to their own chromosomes （called self-genomic in situ hybridization, self-GISH） was carried out in six selected plant species with different genome size and amount of repetitive DNA. Nonuniform distribution of the fluorescent labeled probe DNA was observed on the chromosomes of all the species that were tested. The signal patterns varied among species and were related to the genome size. The chromosomes of the small Arabidopsis genome were labeled almost only in the pericentromeric regions and the nucleolus organizer regions （NORs）. The signals in the relatively small genomes, rice, sorghum, and Brassica oleracea var. capitata L., were dispersed along the chromosome lengths, with a predominant distribution in the pericentromeric or proximal regions and some heterochromatic arms. All chromosomes of the large genomes, maize and barley, were densely labeled with strongly labeled regions and weakly labeled or unlabeled regions being arranged alternatively throughout the lengths. In addition, enhanced signal bands were shown in all pericentromeres and the NORs in B. oleracea var. capitata, and in all pericentromeric regions and certain intercalary sites in barley. The enhanced signal band pattern in barley was found consistent with the N-banding pattern of this species. The GISH with self-genomic DNA was compared with FISH with Cot-1 DNA in rice, and their signal patterns are found to be basically consistent. Our results showed that the self-GISH signals actually reflected the hybridization of genomic repetitive DNAs to the chromosomes, thus the self-GISH technique would be useful for revealing the distribution of the regions where repetitive DNAs concentrate along chromosomes and some chromatin differentiation associated with repetitive DNAs in plants.

A Method for Rapid Salt-extraction of High-quality Genomic DNA from Plant Seeds

[Objective] This study aimed to explore an effective method for rapid salt- extraction of high-quality genomic DNA from dried seeds of plants. [Method] Seeds of seven varieties of crops were ground into powder. A hundred milligrams of seed powder was added to extracting solution for high salt-extraction of genomic DNA. The yield and quality of extracted DNA were determined by using ultramicro UV/Vis spectrophotometer detection method, PCR and restriction enzyme digestion. [Result] About 619.67-1 811.21 ng of genomic DNA was extracted from per 100 mg of dried seed powder of seven varieties of conventional crops. A260/A280 ratios of the obtained DNA solution all ranged from 1.87 to 2.07, the purity and quality of PCR were suitable for PCR and restriction enzyme digestion. Clear target bands of specific endogenous gene fragments of seven varieties of crops were amplified by PCR, and the obtained DNA could be fully digested with EcoRV and Hindlll.[Conclusion] This method could be used for rapid extraction of high-quality genomic DNA from dried seeds.

A New Sampling Approach to Obtain Genomic DNA of Marine Shellfish

[Objective] The aim was to explore a new sampling approach to obtain genomic DNA of marine shellfish,as well as to provide reference for the molecular biology research on precious shellfish.[Method] Meretrix meretrix,Atrina pectinata,Perna viridis,Crassostrea hongkongensis and Scapharca kagoshimensis were used as experimental materials and the genomic DNA of adductor muscle was taken as reference to extract the genomic DNA of shell cavity fluids with the conventional phenol-chloroform method.And then biophotometer,agarose gel electrophoresis,amplification and sequencing of the target fragments were used to examine the quality of genomic DNA.At the same time,the phylogenic tree was constructed to verify the reality of source contributions.[Result] The quality of genomic DNA of shell cavity fluids extracted by the phenol-chloroform method was better than the genomic of adductor muscle.The genomic DNA extracted by this method showed less content of protein,polyphenol and pigment,which could completely meet the demands of amplification and sequencing of the target fragments.Through the phylogenic tree,it was verified that the source contributions of shell cavity fluids were not come from foreign pollutions.[Conclusion] It is completely feasible to obtain the genomic DNA from shell cavity fluids,which could be applied in the target fragments 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.

Studies on the Extraction Methods of Metagenomic DNA from Mud Volcano in Xinjiang

[Objective]To seek one effective extraction method of metagenomic DNA from mud volcano.[Method]The metagenomic DNA from mud volcano was extracted by CTAB extraction method,SDS-enzyme method,improved method,reagent kit method.The extraction of four kinds of methods were compared.[Result]The extracted rate in reagent sets method was the highest,next was improved method,the extracted quantity in SDS-enzyme method was maximum.DNA extracted by the improved method was diluted ten times for PCR.[Conclusion]Considering economy and purity,the improved method can be used as one effective extraction method of metagenomic DNA from mud volcano.

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.

Isolation of Microsatellite DNA and Preliminary Genomic Analysis of Mud Carp (Cirrhina molitorella)

In order to determine the applicability of microsatellite primers developed from common carp （ Cyprinuscario ） for genomic analysis in mud carp （ Cirrhina molitorella ）, 24 primer pairs from common carp were designed to amplify microsatellite loci in the mud carp containing CA, GA, AT and GGGA sequences. Thirteen primers （54%） successfully amplified specific products in the mud carp and 11 primers （48%） showed high polymorphism in the mud carp population. The results indicated that the average number of alleles per locus in the mud carp stocks was 5.2. Average heterozygosity （Ho）, unbiased expected heterozygosity （He） and polymorphism information content （PIC） in the wild population were 0.61 ± 0.2, 0.8 ± 0.09 and 0.72 ± 0.1 respectively. Several Hardy-Weinberg departure value were significandy departed from Hardy-Weinberg equilibrium. The study showed that microsatellite primers from a species of Cyprinidae can be used for mud carp genetic analysis without much cost or time input.

A Rapid and Efficient Method for Preparing Genomic DNA from Microbacterium sp.

A new method was used to preparing genomic DNA from Microbacterium sp.quickly and efficiently.DNA quantity and purity was measured by UV absorbance.Integrity of the genomic DNA was tested by agarose gel eletrophoresis.The DNA prepared by this method was sufficiently pure for PCR.This method saves time and cost,practices easily as well.

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.

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.

Comparison of Different Extraction Methods of Genomic DNA of Raw Soybean Milk

[Objective] The paper was too explore and compare methods of DNA extraction from raw soybean milk.[Method] Taken the soybean milk purchased from market as the material,pyrolysis method,isopropanol precipitation method,CTAB method,SDS method,high-salt low-pH and guanidine isothiocyanate method,as well as their improved methods were used to extract genomic DNA,and the extraction effects of these methods were compared by detecting the DNA using optical density,agarosegel electrophoresis and polymerase chain reaction（PCR）methods.[Result] The genomic DNA extracted by all methods except isopropanol precipitation method could be used in PCR reaction.Meanwhile,the high DNA concentration and purity will be gained by different methods in the order of high-salt low-pH method,high-salt low-pH method,improved CTAB method,improved isopropanol precipitation method,guanidine isothiocyanate method and improved pyrolysis method.[Conclusion] These methods are simply to operate,fast to gain results,and suitable for the extraction of total DNA from raw soybean milk.