Journal of Materials Chemistry B
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2024
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IF 6.1
Xiuli Wang,Qilin Guo,Jia Guo,Changchun Wang
Rapid extraction and screening of high purity DNA fragments is an indispensable technology in advanced molecular biology. In this article, mesoporous magnetic composite microspheres (MSP@mTiO2) with tunable pore sizes are successfully fabricated for high purity DNA extraction and fragment screening. Due to the strong complexation ability of Ti4+ with DNA phosphate groups and the high specific surface area of mesoporous microspheres, the MSP@mTiO2 microspheres possess excellent adsorption performance, the saturated loading capacity of MSP@mTiO2 with specific surface area of 122 m2/g is as high as 575 g/mg for salmon sperm specimen. ITC experiment demonstrates that the DNA adsorption on MSP@mTiO2 microspheres is mainly driven by entropy, which gives us more potential ways to regulate the balance of adsorption and desorption. Meanwhile, the mesoporous MSP@mTiO2 microspheres exhibit much higher extraction efficiency compared with non-porous MSP@TiO2 for whole genome DNA from Arabidopsis thaliana plants. Interestingly, the DNA frangements with different lengths can be screened by simply regulating the pore size of MSP@mTiO2 or concentration of Na3PO4 in eluent. Small pore size and low phosphate concentration are advantageous for the extraction of short-stranded DNA fragments, the DNA fragments (≤1000 bp) can be efficiently extracted when the mesopore size of MSP@mTiO2 is lower than 7.6 nm. The extraction results from the mesoporous composite microspheres provide new promising insights on the DNA purification and screening from complex biological samples.
研究领域:
分子生物学
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材料科学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
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2023
·
IF 9.5
Xiuli Wang,Weiwei Fei,Zhifan Zhou,Mengjing Zhu,Yinghao Chang,Qilin Guo,Jia Guo,Changchun Wang
Magnetic-assisted DNA testing technology has attracted much attention in genetics, clinical diagnostics, environmental microbiology, and molecular biology. However, achieving satisfying DNA adsorption and desorption efficiency in real samples is still a big challenge. In this paper, a new kind of high-quality magnetic composite microsphere of MM@PGMA–PA-Ti4+ was designed and prepared for DNA extraction and detection based on the strong interaction of Ti4+ and phosphate groups. By taking the advantages of high magnetic susceptibility and high Ti4+ content, the MM@PGMA–PA-Ti4+ microspheres possessed remarkable extraction capacity for mimic biological samples (salmon sperm specimens) with saturated loadings up to 533.0 mg/g. When the DNA feeding amount was 100 μg and the MM@PGMA–PA-Ti4+ dosage was 1 mg, the adsorption and desorption efficiencies were 80 and 90%, respectively. The kinetic and equilibrium extraction data were found to fit well with the pseudo-second-order model and Freundlich isotherm model. Furthermore, the MM@PGMA–PA-Ti4+ microspheres were successfully employed for DNA extraction from mouse epithelial-like fibroblasts. The extraction ability (84 ± 4 μg/mg) and DNA purity were superior to the comparative commercial spin kits, as evaluated by electrophoresis assays and qPCR analysis. The experimental results suggest that the MM@PGMA–PA-Ti4+ microspheres possess great potential as an adsorbent for DNA purification from complex biological samples.
研究领域:
分子生物学
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临床诊断学
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环境微生物学
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DOI
PubMed
高影响因子
FOOD CONTROL
·
2025
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IF 6.3
Kai Chen,Bin Xiao,JianZhi Luo,XueWei Yuan,Juan Zhang,Songxue Wang,Rui Zhang,Nan Wang,LiQiang Liu,AiKe Li,Ailiang Chen
Edible oil adulteration is a significant food safety concern in today's food fraud landscape. DNA-based detection methods emerging as the most reliable approach for identifying adulterated products. However, the processed oil matrix contains only trace amounts of highly degraded DNA, which significantly increases the difficulty of DNA extraction and greatly reduces the success rate of PCR amplification. To address these technical challenges, we designed species-specific primers for amplicons of varying lengths and systematically evaluated their performance in real-time PCR using DNA extracted from soybean, peanut, and rapeseed oils, followed by application in adulterated oil detection. All primer sets demonstrated high specificity, sensitivity and, most importantly, equal amplification efficiency. Results indicated that shorter amplicons were consistently detectable across all three vegetable oils, while longer amplicons were more likely to yield false negatives in real-time PCR reactions. Soybean oil showed CT values of 29 (74 bp) versus 38 (170 bp), peanut oil exhibited a CT value of 31 (54 bp) with no amplification for 86 bp, and rapeseed oil displayed a CT value of 34 (66 bp) with no signal for 101 bp. The use of shorter primers successfully facilitated the detection of adulterated oil.These findings suggest that amplicon length being a critical determinant for reliable DNA-based identification and authentication of edible oils. Thus, designing amplicons within the 50–80 bp range can significantly enhance the success rate of DNA-based identification and authentication of edible oils.
研究领域:
食品科学
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分子生物学
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DOI
高影响因子
MICROCHIMICA ACTA
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2017
·Vol. 184
·期 10
·4133-4140
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IF 4.58
Xu Kaijia,Wang Yuzhi,Zhang Hongmei,Yang Qin,Wei Xiaoxiao,Xu Panli,Zhou Yigang
A magnetically responsive composite was prepared for improved solid-phase extraction of salmon sperm DNA sodium salt. It consists of chitosan-modified Fe 3 O 4 -magnetized multi-walled carbon nanotubes coated with a poly(ethylene glycol) based deep eutectic solvent. The sorbent was characterized by X-ray diffraction, vibrating sample magnetometry, Fourier transform infrared spectrometry, thermogravimetric analysis and transmission electron microscopy. The effects of the concentration of DNA, ionic strength, pH value, temperature and extraction time on extraction performance were optimized. Compared to plain Fe 3 O 4 , the new sorbent displays a superior DNA extraction capacity of 178 mg·g −1 . It also displays favorable selectivity for DNA over bovine hemoglobin. Regeneration studies show that (a) 79% of DNA can be eluted from loaded sorbent by using 1 M NaCl, and that (b) the conformation of DNA remains unchanged as evidenced by CD spectra. The sorbent can be recycled six times without significant loss of extraction capacity. It was applied to the extraction of DNA from bovine whole blood prior to DNA quantitation by agarose gel electrophoresis. The results show this new sorbent to be a viable material for separation of DNA that excels by its low cost, high loading capacity and ease of regeneration.
Chitosan (CS)-modified magnetic multi-walled carbon nanotubes were coated with a poly(ethylene glycol)-based deep eutectic solvent (DES) to obtain a new material (referred to as DES-mCS/MWCNTs) for solid-phase extraction of DNA. It exhibits excellent magnetic response, high loading capacity and good reusability.
研究领域:
生物技术
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材料科学
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DOI
高影响因子
CHEMICAL PHYSICS LETTERS
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2018
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IF 1.686
Qingqian Fan,Yueping Guan,Zhi Zhang,Guoli Xu,Yu Yang,Chen Guo
A new method was developed for preparing monodisperse Fe 3 O 4 @SiO 2 magnetic core-shell nanoparticles via three steps. The obtained nanoparticles exhibit high saturation magnetization, good surface topography and dispersion as characterized by FTIR, SEM and VSM. The experimental results of hydrochloric acid treatment show that the obtained nanoparticles have excellent acid resistance and stable and compact silicon coating. The magnetic core-shell nanoparticles were further tested for purification of plasmid DNA from E. coli DH5a cells. The result of purification leaded to better adsorption, elution and purity results in comparison with isolated commercial extraction kit.
研究领域:
生物技术
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材料科学
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分子生物学
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DOI
高影响因子
ACS Applied Materials & Interfaces
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2023
·
IF 9.5
Zhongjie Fei,Ping Liu,Chu Cheng,Rongbin Wei,Pengfeng Xiao,Youfa Zhang
Although numerous approaches were proposed for the nucleic acid (NA)-based SARS-CoV-2 detection, the nonideal NA desorption efficiency of conventional magnetic beads (MBs) limits their widespread application. In this study, we developed solvent-responsive MBs (called responsive MBs), which, in the presence of buffers, modulated the absorption and desorption capacities of NA by flipping the surface −COO–. Relative to other commercial MBs, responsive MBs exhibited similar absorption profiles and markedly enhanced desorption profiles. When applied for NA detection of complex samples, responsive MBs exhibited better performance of RNA detection than DNA, with obvious advantages in sensitivity. Specifically, the RNA and DNA desorption rates of commercial MBs were ∼85 and 82.5%, while those of responsive MBs were nearly 94 and 93.5%, respectively. Furthermore, responsive MBs exhibited remarkable extraction ability in a wide range of tissues and better performance of RNA extraction than DNA. When applied for SARS-CoV-2 detection, the responsive MBs along with the simulated digital RT-LAMP (a previously established apparatus) further improved detection efficiency, yielding a precise quantitative detection as low as 25 copies and an ultimate sensibility detection of 5 copies/mL. It was also successfully employed in numerous NA-based technologies such as polymerase chain reaction (PCR), sequencing, and so on.
研究领域:
生物技术
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分子生物学
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病毒学
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分析化学
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DOI
PubMed
高影响因子
JOURNAL OF MATERIALS RESEARCH
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2012
·Vol. 27
·期 7
·1006-1013
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IF 1.434
Guoxin Zhang,Fengxiang Qie,Jianxuan Hou,Shizhong Luo,Liang Luo,Xiaoming Sun,Tianwei Tan
Surface-functionalized magnetic nanoparticles were prepared by a facile one-pot solvothermal method in ethylene glycol solution. Zeta value, size, and magnetic properties could be well tuned by introducing different functional group molecules. Characterizations, including transmission electronic microscopy, scanning electronic microscopy, thermogravimetric analysis, x-ray powder diffraction and vibrating sample magnetometer, and Fourier transform infrared spectrophotometer demonstrated the efficiency of this simple and general synthesis strategy. The hydrophilic magnetic nanoparticles with various surface functional groups and zeta values were evidenced as excellent candidates for bioseparation by extracting DNA molecules from a model mixture of cell fractures.
研究领域:
纳米医学
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生物材料
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分子生物学
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生物分离技术
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纳米生物技术
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DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2024
·Vol. 25
·期 24
·13599
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IF 4.9
Haibin Liu,Xinyue Wang,Hang Tian,Yi Yuan,Jing Wang,Yani Cheng,Linyao Sun,Hongshuo Chen,Xiaoming Song
Due to the price and demand ofOphiocordyceps sinensishaving increased dramatically, adulteration with other fungi is a common problem. Thus, a reliable method of authenticO. sinensisidentification is essential. In the present work, a rapid DNA extraction and double-tailed recombinase polymerase amplification (RPA) coupled with nucleic acid hybridization lateral flow strip (NAH-LFS) was developed to distinguish authenticO. sinensisingredients from other fungi substitutes. In the presence ofO. sinensis, the RPA amplicons with two ssDNA tails in the opposite ends, which could simultaneously bind with the SH-probes on gold nanoparticles (AuNPs) and capture the probe on the test line, formed visible red bands. RPA combined with NAH-LFS can efficiently detectO. sinensisDNA down to 1.4 ng/μL; meanwhile, the specificity test validated no cross reaction with common adulterants, includingCordyceps gunnii,Cordyceps cicadae,Cordyceps militaris,yungui Cordyceps, andOphiocordyceps nutans. The whole RPA-NAH-LFS could be completed within 16 min. The RPA-NAH-LFS results in detecting 20 commercialO. sinensissamples are consistent with PCR-AGE and RT-PCR, confirming the feasibility of the RPA-NAH-LFS method. In conclusion, these results are expected to facilitate the application of RPA-NAH-LFS in the authentication detection ofO. sinensismaterials, providing a convenient and efficient method forO. sinensisquality control.
研究领域:
分子生物学
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中药学
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生物技术
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PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2021
·
IF 5.279
Huan Li,Jiesi Feng,Yanbo Wang,Guangming Liu,Xiaojing Chen,Linglin Fu
DNA-based detection methods are highly promising for risk assessment in the food sector, such as tracing the existence of food allergens. However, due to the complexity of food matrices, cumbersome protocols are often needed to isolate the DNA components, which hinder the achievement of rapid and on-site detection. Herein, an instant and multiple DNA extraction method was developed based on the poly(vinyl alcohol) microneedle (MN) patch. With simple press and peel-off operations within 1 min, samples suitable for DNA-based analysis such as polymerase chain reaction (PCR) could be collected. By further combining with the recombinase polymerase amplification assay, rapid screening of the allergenic risks in complex samples such as shrimp ball and cheesecake could be achieved within 30 min. The MN-based DNA extraction method not only was a potential alternative to the traditional DNA extraction method but provided a transformative approach in realizing rapid, on-site detection of foodborne hazards in collaborating with fast DNA-based assays.
研究领域:
食品科学
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生物技术
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分子生物学
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DOI
PubMed
高影响因子
RSC Advances
·
2023
·Vol. 13
·期 14
·9595-9602
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IF 3.9
Jia Xu,Yuan Yang,Xiaonan Cai,Han Xiao
In this study, hexafluoroisopropanol (HFIP)-based deep eutectic solvents (DESs) were developed and used for DNA extraction from human whole blood samples for the first time. HFIP-based DESs were prepared using HFIP and choline chloride (ChCl)/tetrabutylammonium chloride/cetyltrimethylammonium bromide as the hydrogen bond donor and acceptor, respectively. The two-phase system formation was promoted with different inorganic salts as the phase-forming component. According to the strong phase separation capability and high DNA extraction efficiency, DESs consisting of HFIP/ChCl-(NH4)2SO4, HFIP/ChCl-Na2SO4 and HFIP/ChCl-MgSO4 were then selected for application in DNA extraction. The factors that could have impacted the DNA extraction process, including molar ratio of DES, DES addition, salt species and addition, and sample pH, were systematically investigated via single-factor experimental analysis. Furthermore, we selectively examined bovine serum albumin and RNA to assess the specificity of the HFIP-based DESs for DNA extraction. Conclusively, 93.9%, 96.7% and 99.8% DNA could be extracted using the HFIP/ChCl-(NH4)2SO4, HFIP/ChCl-Na2SO4, and HFIP/ChCl-MgSO4 systems, respectively. Moreover, the developed systems were successfully used to extract DNA from human whole blood with satisfactory results. The DNA secondary structure was stable after DES extraction with the electrostatic interaction between DES and DNA as the main force driving DNA adsorption by DES.
研究领域:
化学
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生物化学
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分子生物学
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分析化学
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DOI
PubMed
高影响因子
ACS Nano
·
2023
·
IF 17.1
Cuiping Mao,Shuchen Wang,Jiankai Li,Zhuowei Feng,Tong Zhang,Rui Wang,Chunhai Fan,Xingyu Jiang
DNA as an exceptional data storage medium offers high information density. However, DNA storage requires specialized equipment and tightly controlled environments for storage. Fast encapsulation within minutes for enhanced DNA stability to do away with specialized equipment and fast DNA extraction remain a challenge. Here, we report a DNA microlibrary that can be encapsulated by metal–organic frameworks (MOFs) within 10 min and extracted (5 min) in a single microfluidic chip for automated and integrated DNA-based data storage. The DNA microlibrary@MOFs enhances the stability of data-encoded DNA against harsh environments. The encoded information can be read out perfectly after accelerated aging, equivalent to being readable after 10 years of storage at 25 °C, 50% relative humidity, and 10 000 lx sunlight radiation. Moreover, the library enables fast retrieval of target data via flow cytometry and can be reproduced after each access.
研究领域:
材料科学
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生物技术
·
数据存储技术
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化学工程
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2023
·
IF 8.8
Bin Xiao,Ruiming Zhao,Nan Wang,Juan Zhang,Xiaoyun Sun,Fengchun Huang,Ailiang Chen
Most microfluidic-based “sample-in-result-out” systems suffer sophisticated microfluidic production processes, high-cost chips, and expensive instruments. They cannot be used in the meat market as well as farmer's markets in rural areas. Here, we developed a hand-held microfluidic chip system for on-site meat species qualitative authentication detection which integrated a simple microneedle DNA extraction and a visual loop-mediated isothermal amplification (LAMP). The chip can be used by easily pricking meat samples, simply hand-shaking the chip, and readily available isothermal heating instead of a complicated DNA extraction process and microfluidic control device. The system demonstrates high specificity and sensitivity for selected six species of meat samples and low to 1% simulated adulteration could be detected within 60 min. Besides, the whole cost was less than 1 dollar. The integrated hand-held microfluidic detection system offers a simple, fast, low-cost “sample-in-result-out” point-of-care device which could be extended to medical diagnosis and animal/plant disease identification.
研究领域:
生物技术
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食品科学
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分子生物学
·
即时诊断
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）