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

被引：141

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

来源：

SCIENCE CHINA-CHEMISTRY | 2015年 / 58卷 / 11期

基金：

中国国家自然科学基金; 国家高技术研究发展计划(863计划); 中国博士后科学基金;

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

页码：1774 / 1778

页数：5

共 44 条

[1] RNA interference with DNA polymerase and synthesis [J].