Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles

被引:139
|
作者
Wang, Jiuhai [1 ]
Ali, Zeeshan [1 ]
Wang, Nianyue [2 ]
Liang, Wenbiao [3 ]
Liu, Hongna [1 ]
Li, Fu [1 ]
Yang, Haowen [1 ]
He, Lei [1 ]
Nie, Libo [4 ]
He, Nongyue [1 ,4 ]
Li, Zhiyang [5 ]
机构
[1] Southeast Univ, Sch Biol Sci & Med Engn, State Key Lab Bioelect, Nanjing 210096, Jiangsu, Peoples R China
[2] Second Hosp Nanjing, Clin Lab, Nanjing 210003, Jiangsu, Peoples R China
[3] Jiangsu Prov Blood Ctr, Nanjing 210042, Jiangsu, Peoples R China
[4] Hunan Univ Technol, Econ Forest Cultivat & Utilizat Collaborat Innova, Hunan Key Lab Green Packaging & Applicat Biol Nan, Zhuzhou 412007, Peoples R China
[5] Nanjing Med Univ, Affiliated Hosp 2, Dept Lab Med, Nanjing 210011, Jiangsu, Peoples R China
基金
国家高技术研究发展计划(863计划); 中国博士后科学基金; 中国国家自然科学基金;
关键词
nucleic acids; simultaneous extraction; magnetic nanoparticles (MNPs); bacteria; NUCLEIC-ACIDS; GENOMIC DNA; ADSORPTION; SEPARATION; PARTICLES;
D O I
10.1007/s11426-015-5483-x
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles (MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease (DNase) and Ribonucleases (RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.
引用
收藏
页码:1774 / 1778
页数:5
相关论文
共 50 条
  • [31] Phenyl-functionalized silica-coated magnetic nanoparticles for the extraction of chlorobenzenes, and their determination by GC-electron capture detection
    Saraji, Mohammad
    Khaje, Nasrin
    JOURNAL OF SEPARATION SCIENCE, 2013, 36 (06) : 1090 - 1096
  • [32] Immobilization of α-transglucosidase on silica-coated magnetic nanoparticles and its application for production of isomaltooligosaccharide from the potato peel
    Maurya, Rohit
    Ali, Usman
    Kaul, Sunaina
    Bhaiyya, Raja
    Singh, Ravindra Pal
    Mazumder, Koushik
    SCIENTIFIC REPORTS, 2023, 13 (01):
  • [33] Immobilization of α-transglucosidase on silica-coated magnetic nanoparticles and its application for production of isomaltooligosaccharide from the potato peel
    Rohit Maurya
    Usman Ali
    Sunaina Kaul
    Raja Bhaiyya
    Ravindra Pal Singh
    Koushik Mazumder
    Scientific Reports, 13 (1)
  • [34] Ionic liquid-modified silica-coated magnetic nanoparticles: promising adsorbents for ultra-fast extraction of paraquat from aqueous solution
    Latifeh, Farzad
    Yamini, Yadollah
    Seidi, Shahram
    ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH, 2016, 23 (05) : 4411 - 4421
  • [35] Ionic liquid-modified silica-coated magnetic nanoparticles: promising adsorbents for ultra-fast extraction of paraquat from aqueous solution
    Farzad Latifeh
    Yadollah Yamini
    Shahram Seidi
    Environmental Science and Pollution Research, 2016, 23 : 4411 - 4421
  • [36] Silica-coated magnetic beads containing an Fe-based amorphous alloy are applicable for nucleic acid extraction
    Fukuchi, Maika
    Hanamura, Masato
    Otake, Kichiya
    Nakamori, Michio
    Toyoda, Naoyuki
    Matsuda, Kazuyuki
    JOURNAL OF NON-CRYSTALLINE SOLIDS, 2023, 609
  • [37] Molybdenum (VI) complex of resorcinol-based ligand immobilized on silica-coated magnetic nanoparticles for biodiesel production
    Majedi, Mona
    Safaei, Elham
    APPLIED ORGANOMETALLIC CHEMISTRY, 2023, 37 (10)
  • [38] Ionic liquid-modified silica-coated magnetic nanoparticles; promising anion-exchange sorbent for extraction of Cr(VI)
    Yamini, Yadollah
    Seidi, Shahram
    Latifeh, Farzad
    INTERNATIONAL JOURNAL OF ENVIRONMENTAL ANALYTICAL CHEMISTRY, 2017, 97 (13) : 1223 - 1236
  • [39] Self-cleaning superhydrophobic epoxy coating based on fibrous silica-coated iron oxide magnetic nanoparticles
    Alamri, Haleema
    Al-Shahrani, Abdullah
    Bovero, Enrico
    Khaldi, Turki
    Alabedi, Gasan
    Obaid, Waleed
    Al-Taie, Ihsan
    Fihri, Aziz
    JOURNAL OF COLLOID AND INTERFACE SCIENCE, 2018, 513 : 349 - 356
  • [40] Functionalization of silica-coated magnetic nanoparticles as powerful demulsifier to recover oil from oil-in-water emulsion
    Elmobarak, Wamda Faisal
    Almomani, Fares
    CHEMOSPHERE, 2021, 279