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 comparison study on structure-function relationship of polysaccharides obtained from sea buckthorn berries using different methods:antioxidant and bile acid-binding capacity

In this study,the structural characters,antioxidant activities and bile acid-binding ability of sea buckthorn polysaccharides(HRPs)obtained by the commonly used hot water(HRP-W),pressurized hot water(HRP-H),ultrasonic(HRP-U),acid(HRP-C)and alkali(HRP-A)assisted extraction methods were investigated.The results demonstrated that extraction methods had significant effects on extraction yield,monosaccharide composition,molecular weight,particle size,triple-helical structure,and surface morphology of HRPs except for the major linkage bands.Thermogravimetric analysis showed that HRP-U with filamentous reticular microstructure exhibited better thermal stability.The HRP-A with the lowest molecular weight and highest arabinose content possessed the best antioxidant activities.Moreover,the rheological analysis indicated that HRPs with higher galacturonic acid content and molecular weight showed higher viscosity and stronger crosslinking network(HRP-C,HRP-W and HRP-U),which exhibited stronger bile acid binding capacity.The present findings provide scientific evidence in the preparation technology of sea buckthorn polysaccharides with good antioxidant and bile acid binding capacity which are related to the structure affected by the extraction methods.

A Review on Sources,Extractions and Analysis Methods of a Sustainable Biomaterial:Tannins

Condensed and hydrolysable tannins are non-toxic natural polyphenols that are a commercial commodity industrialized for tanning hides to obtain leather and for a growing number of other industrial applications mainly to substitute petroleum-based products.They are a definite class of sustainable materials of the forestry industry.They have been in operation for hundreds of years to manufacture leather and now for a growing number of applications in a variety of other industries,such as wood adhesives,metal coating,pharmaceutical/medical applications and several others.This review presents the main sources,either already or potentially commercial of this forestry by-materials,their industrial and laboratory extraction systems,their systems of analysis with their advantages and drawbacks,be these methods so simple to even appear primitive but nonetheless of proven effectiveness,or very modern and instrumental.It constitutes a basic but essential summary of what is necessary to know of these sustainable materials.In doing so,the review highlights some of the main challenges that remain to be addressed to deliver the quality and economics of tannin supply necessary to fulfill the industrial production requirements for some materials-based uses.

Online identification and extraction method of regional large-scale adjustable load-aggregation characteristics

This article introduces the concept of load aggregation,which involves a comprehensive analysis of loads to acquire their external characteristics for the purpose of modeling and analyzing power systems.The online identification method is a computer-involved approach for data collection,processing,and system identification,commonly used for adaptive control and prediction.This paper proposes a method for dynamically aggregating large-scale adjustable loads to support high proportions of new energy integration,aiming to study the aggregation characteristics of regional large-scale adjustable loads using online identification techniques and feature extraction methods.The experiment selected 300 central air conditioners as the research subject and analyzed their regulation characteristics,economic efficiency,and comfort.The experimental results show that as the adjustment time of the air conditioner increases from 5 minutes to 35 minutes,the stable adjustment quantity during the adjustment period decreases from 28.46 to 3.57,indicating that air conditioning loads can be controlled over a long period and have better adjustment effects in the short term.Overall,the experimental results of this paper demonstrate that analyzing the aggregation characteristics of regional large-scale adjustable loads using online identification techniques and feature extraction algorithms is effective.

Physicochemical properties and biological activities of polysaccharides from the peel of Dioscorea opposita Thunb.extracted by four different methods

Polysaccharides are the important biologically active components found in the peel of Dioscorea opposita Thunb.(DTTP).The influences of 4 extraction methods,namely hot water extraction(W),acidic extraction(HA),hot-compressed water extraction(HCW)and enzyme-assisted extraction(EAE),on the yields,physicochemical properties,hypoglycemic and antioxidant activities of polysaccharides from DTTP were studied and compared.Among these DTTP polysaccharides,DTTP-HA was outstanding in several respects.DTTP-HA was the most water soluble;it had the highest total carbohydrate content(85.08%),the highest uronic acid(13.20%)and the highest thermal stability.DTTP-HA and DTTP-W possessed a triple-helix structure.These 4 kinds of polysaccharides have the same types of monosaccharides,but in different molar percentages.Extraction method had a significant impact on the microstructures of the extracted polysaccharides.DTT-HA exhibited irregular structure with many holes.Among the 4 extracted methods,the DTTP-HA and DTTP-W initially exhibited higher hypoglycemic and antioxidant activities.The better bioactivities of DTTP-HA may be related to the above factors.The findings indicated that acid extraction is an effective method to extract polysaccharides with high biological activities from DTTP.

Effects of dietary oil sources and fat extraction methods on apparent and standardized ileal digestibility of fat and fatty acids in growing pigs

Background:There is a lack of data for the standardized ileal digestibility(SID) of fat and fatty acids in national feed databases.In addition,it is important to specify the procedures used for fat analyses.Therefore,an experiment was conducted to 1) determine the apparent ileal digestibility(AID) and SID of fat and fatty acids in ten different oil sources for growing pigs and to develop prediction equations for SID of fat based on fatty acid composition;and 2) compare the effect of the fat extraction methods on the calculated values for endogenous loss and digestibility of fat.Methods:Twenty-two barrows(initial body weight:32.1 ± 2.3 kg) were surgically fitted with a T-cannula in the distal ileum,and allotted to 1 of 11 experimental diets in a 4-period Youden Square design.A fat-free diet was formulated using cornstarch,soy protein isolate and sucrose.Ten oil-added diets were formulated by adding 6% of dietary oil sources to the fat-free diet at the expense of cornstarch.All diets contained 26% sugar beet pulp and 0.40% chromic oxide.Results:The endogenous loss of ether extract(EE) was lower than that of acid-hydrolyzed fat(AEE;P < 0.01).There were significant differences in the AID and SID of fat and saturated fatty acids across the dietary oil sources(P < 0.05).The SID of AEE for palm oil was lower than that of sunflower oil,corn oil,canola oil,rice oil and flaxseed oil(P < 0.01).The AID and SID of fat ranged from 79.65% to 86.97% and from 91.14% to 99.18%.Although the AID of EE was greater than that of AEE(P < 0.01),there was no significant difference in SID of EE and AEE except for palm oil.The ratio of unsaturated to saturated fatty acids(U/S) had a positive correlation with SID of fat(P < 0.05),whereas C16:0 and long chain saturated fatty acids(LSFA) were significant negatively correlated with SID of fat(P < 0.01).The best-fit equation to predict SID of fat was SID AEE = 102.75-0.15 × LSFA-0.74 × C18:0-0.03 × C18:1(Adjusted coefficient of determination = 0.88,P < 0.01).Conclusions:When calculating the SID of fat,the EE content of the samples can be analyzed using the direct extraction method,whereas the acid hydrolysis procedure should be used to determine the AID of fat.Fat digestibility of dietary oils was affected by their fatty acid composition,especially by the contents of C16:0,LSFA and U/S.

Comparison of Three Methods in Extracting Coherent Modes from a Doppler Backscatter System

We compare three different methods to extract coherent modes from Doppler backscattering （DBS）, which are center of gravity （COG） of the complex amplitude spectrum, spectrum of DBS phase derivative （phase derivative method）, and phase spectrum, respectively. These three methods are all feasible to extract coherent modes, for example, geodesic acoustic mode oscillation. However, there are stilI differences between dealing with high frequency modes （several hundred kHz） and low frequency modes, （several kHz） hiding in DBS signal. There is a significant amount of power at low frequencies in the phase spectrum, which can be removed by using the phase derivative method and COG. High frequency modes are clearer by using the COO and the phase derivative method than the phase spectrum. The spectrum of DBS amplitude does not show the coherent modes detected by using COG, phase derivative method and phase spectrum. When two Doppler shifted peaks exist, coherent modes and their harmonics appear in the spectrum of DBS amplitude, which are introduced by the DBS phase.

Robotic natural orifice specimen extraction surgery I-type F method vs conventional robotic resection for lower rectal cancer

BACKGROUND Robotic resection using the natural orifice specimen extraction surgery I-type F method(R-NOSES I-F)is a novel minimally invasive surgical strategy for the treatment of lower rectal cancer.However,the current literature on this method is limited to case reports,and further investigation into its safety and feasibility is warranted.AIM To evaluate the safety and feasibility of R-NOSES I-F for the treatment of low rectal cancer.METHODS From September 2018 to February 2022,206 patients diagnosed with low rectal cancer at First Affiliated Hospital of Nanchang University were included in this retrospective analysis.Of these patients,22 underwent R-NOSES I-F surgery(RNOSES I-F group)and 76 underwent conventional robotic-assisted low rectal cancer resection(RLRC group).Clinicopathological data of all patients were collected and analyzed.Postoperative outcomes and prognoses were compared between the two groups.Statistical analysis was performed using SPSS software.RESULTS Patients in the R-NOSES I-F group had a significantly lower visual analog score for pain on postoperative day 1(1.7±0.7 vs 2.2±0.6,P=0.003)and shorter postoperative anal venting time(2.7±0.6 vs 3.5±0.7,P<0.001)than those in the RLRC group.There were no significant differences between the two groups in terms of sex,age,body mass index,tumor size,TNM stage,operative time,intrao-perative bleeding,postoperative complications,or inflammatory response(P>0.05).Postoperative anal and urinary functions,as assessed by Wexner,low anterior resection syndrome,and International Prostate Symptom Scale scores,were similar in both groups(P>0.05).Long-term follow-up revealed no significant differences in the rates of local recurrence and distant metastasis between the two groups(P>0.05).CONCLUSION R-NOSES I-F is a safe and effective minimally invasive procedure for the treatment of lower rectal cancer.It improves pain relief,promotes gastrointestinal function recovery,and helps avoid incision-related complications.

Optimization of extraction method for quantitative analysis of Si/Al in soil phytoliths

Elemental information carried by phytoliths plays a crucial indicative role in geochemical research.For instance,it serves as an indicator of the carbon pool effect in phytoliths and aids in the elucidation of silicon sources.However,early extraction methods for soil phytoliths primarily focused on obtaining their morphological and quantitative information,lacking efficient techniques for quantitative elemental analysis.In this study,we aimed to extract Si/Al information from soil phytoliths.Considering the need for complete extraction of phytolith,six extraction methods were developed and further by alkaline dissolution to determine Si/Al.Six methods were compared in terms of enrichment capacity,the weight of extracted phytoliths,and Si/Al differences.The results indicated that the addition of Ammonia-Catechol in the commonly used heavy liquid flotation method effectively improved phytolith extraction capability and the accuracy of Si/Al results.Additionally,the inclusion of an acetic acid step before alkaline dissolution further removed surface-adsorbed impurities and enhanced the analytical quality of Si/Al in phytolith.The comparison of the data in this study with other published data shows that our method is relatively robust.The improved method proposed in this study can provide a new idea for the quantitative analysis of other elements in soil phytoliths.

Microplastics in Marine Environment: Occurrence, Distribution, and Extraction Methods in Marine Organisms

The pervasive presence of microplastics in marine environments has raised significant concerns. This review addresses the pressing issue of microplastic pollution in marine ecosystems and its potential implications for both the environment and human health. It outlines the current state of microplastic occurrence, distribution, and extraction methods within marine organisms. Microplastics have emerged as a significant environmental concern due to their harmful effects on ecosystems and their potential human health risks. These particles infiltrate marine environments through runoff and atmospheric deposition, ultimately contaminating beaches and posing threats to marine life. Despite the gravity of this issue, there has been limited research on the presence and distribution of microplastics in marine organisms. This review aims to bridge this knowledge gap by comprehensively examining the occurrence, distribution, and various extraction methods used to detect microplastics in marine organisms. It emphasizes the urgent need for targeted measures to manage microplastic pollution, highlights the significant role of human activities in contributing to this problem, and underscores the importance of reducing human-induced pollution to safeguard marine ecosystems. While this paper contributes to the understanding of microplastic pollution in marine environments and underscores the critical importance of taking action to protect marine organisms and preserve our oceans for future generations, it also emphasizes that, in effectively tackling the microplastic problem, a well-coordinated approach is essential, involving research initiatives, policy adjustments, public involvement, and innovative technologies. Crucially, prompt and resolute responses must exist to counteract the escalating peril posed by microplastics to the oceans and the global environment.

Comparison of RNA Extraction Methods of Sugarcane Stem

[Objective] The study was to explore the most effective and feasible method for sugarcane stalks RNA extraction.[Method] SDS extraction method,kit extraction method and GHCL extraction method were used to extract the sugarcane RNA in stalks,and the quality of the extracted RNA was compared.[Result] RNA extracted by kit extraction method had a high-yield,the bands were clear and RNA had a good integrity,there was no significant degradation of RNA,and the OD260 nm/OD280 nm value was closed to 2.0.[Conclusion] Kit extraction method was the effectively method to extract sugarcane RNA,and this study had provided a theoretical basis for the molecular biology study of sugarcane.

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.

Extraction of Pumpkin Polysaccharide by Complex Enzyme Method and Its Antioxidant Research

[Objective] This paper aims to study a new method of extracting pumpkin polysaccharide from pumpkin. Single factor experiments were conducted to examine the effects of extracting time,temperature,the solid-liquid ratio and pH value on the extraction yield of polysaccharide from pumpkin. [Method] The best enzyme ratio and extraction conditions for complex enzymes extraction were determined through orthogonal tests. Scavenging ·OH and O-2 activities of pumpkin polysaccharides were also investigated by salicylic acid and improved self-oxidation of o-pheno methods respectively. [Results] The results showed that the biggest extraction yield of polysaccharide from pumpkin can be got when adding 1% cellulose enzyme,1.5% pectinase,1.0% papain and Na2HPO4-citric acid buffer solution (pH was 4.6),and oscillating for 30 min under water at 40 ℃ with the solid-liquid ratio of 1:30. In addition,pumpkin polysaccharides had a strong activity of eliminating ·OH,but very weak activity to scavenge O-2. [Conclusion] This study provided basic data for research and application of Pumpkin polysaccharide.

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.

Progress in the extraction and application of collagen peptides

Collagen peptide is the product of complete hydrolysis of collagen,which has a relatively small molecular weight and is more easily absorbed than proteins and amino acids.Collagen peptide not only has unique nutritional value,but also has certain physiological functions,which makes it has great potential value in various fields,so it has set off a wave of research on collagen peptide in the biological world.This paper describes the sources and extraction methods of collagen peptides,and describes the research progress and application of collagen peptides in the medical,food,material and skin care industries according to their physiological functions,which will provide new ideas for the future research of collagen peptides.

A Comparative Study on Three Methods for the Extraction of Total RNA from Pinus bungeana

[Objective] The aim of this study was to compare three RNA extraction methods and thus find out the suitable one for isolating intact and high quality total RNA from Pinus bungeana.[Method] Employing three extraction methods of Trizol,RNeasy Mini Kit,LiCl precipitation,total RNAs of P.bungeana were extracted from pine leaf samples,and their integrity and purity were detected via agarose gel electrophoresis and spectrophotometry for a comparative study.[Result] Among the three extraction methods,LiCl precipitation method demonstrated higher yield and better integrity of total extracted RNA,with OD260/OD280 ratio between 1.8-2.0 and clear 28 S and 18 S bands in electrophoresis pattern.[Conclusion] LiCl precipitation method could be used to extract highly pure and intact total RNA from P.bungeana.

DNA extraction of birch leaves by improved CTAB method and optimization of its ISSR system

The basic method of DNA extraction （CTAB） was improved as the multi-times STE-CTAB extraction method and used to extract the DNA of birch leaved in this experiment. Results showed that the improved method is suitable not only for genomic DNA extraction of birch but also for that of other plants. The purity of genornic DNA extracted by the.multi-times STE-CTAB extraction method is higher than that by one time STE-CTAB method, and it does not need the process of RNase. The factors of influencing ISSR system were explored based on the genomic DNA of birch extracted by the two methods. The optimal conditions for ISSR system were determined as follows： Mg2＋ concentration is 1.5-3.0 mmol·L^-1, dNTP concentration 0.104）.25 mmol·L^-1, the quantity of Taq polymerase 0.5-2.0 U, template DNA 30-100 ng, and the concentration of primer is 0.2-0.4 pmmol·L^-1, and the reaction program was as： initial denaturation for 5 min at 94℃, 30 cycles of denaturation for 30 s at 94℃,annealing for 30 s at 51℃, extension for 30 s at 72℃, and a final 7 min extension at 72℃.

Evaluation Indicators and Extraction Method for Pitting Corrosion of Structural Steel

A set of evaluation indicators based on corrosion ratio in theory for assessing the extent of pitting corrosion and performance reduction are proposed. In order to quantify the morphology of pitting corrosion and extract the evaluation indicators,the 3D profile data obtained by pitting morphology measurement are imported into a special written program to automatically determine the location of each corrosion pit and distill any desired data pertinent to the pitting morphology. The results show that this method seems to be effective to analyze the corroded surface and characterize the pitting morphology.

Study on Extraction Method of Carotenoids from Citrus Leaves

In order to explore the extraction method of carotenoids from citrus leaves,the extraction effect of carotenoids from leaves of Honey pomelo by ultrasonic-assisted extraction was investigated by single factor experiment and orthogonal experiment.The results showed that the optimum extraction condition was:the ration of ethanol and acetone 1∶1(v/v,containing 0.1%BHT),solid-liquid ratio 1∶30(g/mL),extraction time 7 min,ultrasonic power 240 W,extraction frequency 3 times.Under the above condition,the extraction rate of carotenoids was 85.98%.