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学科主题: Chemistry ; Analytical
题名: Extraction of genomic DNA using a new amino silica monolithic column
作者: Liu, Lijia1; Yu, Shengbing1; Yang, Shuixian1; Zhou, Ping1; Hu, Jiming1; Zhang, Yibing2
通讯作者: Zhou, P, Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China
关键词: DNA purification ; Genomic DNA ; Monolithic column ; Solid-phase extraction
刊名: JOURNAL OF SEPARATION SCIENCE
发表日期: 2009-08-01
DOI: 10.1002/jssc.200900208
卷: 32, 期:15-16, 页:2752-2758
收录类别: SCI
文章类型: Article
部门归属: [Liu, Lijia; Yu, Shengbing; Yang, Shuixian; Zhou, Ping; Hu, Jiming] Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China; [Zhang, Yibing] Chinese Acad Sci, Inst Hydrobiol, Wuhan, Peoples R China
WOS标题词: Science & Technology ; Physical Sciences
资助者: Natural Science Foundation of China [20875073]
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
摘要: A new amino silica monolithic column was developed for DNA extraction in a miniaturized format. The monolithic column was prepared in situ by polymerization of tetraethoxysilane (TEOS) and N-(beta-aminoethyl)-gamma-aminopropylmethyldimethoxysilane (AEAPMDMS). DNA was loaded in 50 mM tris(hydroxylmethyl)aminomethane-EDTA buffer at pH 7.0 and eluted with 300 mM potassium phosphate solution at pH 10.0. Under optimal condition, a 6.0-cm monolithic column provided a capacity of 56 ng DNA with an extraction efficiency of 71 +/- 5.2% (X +/- RSD). When the amino silica monolithic column was applied to extract genomic DNA from the whole blood of crucian carp, an extraction efficiency of 52 +/- 5.6% (X +/- SD) was obtained by three extractions. Since the chaotropic-based sample loading and organic solvent wash steps were avoided in this procedure, the purified DNA was suitable for downstream processes such as PCR. This amino silica monolithic column was demonstrated to allow rapid and efficient DNA purification in microscale.
英文摘要: A new amino silica monolithic column was developed for DNA extraction in a miniaturized format. The monolithic column was prepared in situ by polymerization of tetraethoxysilane (TEOS) and N-(beta-aminoethyl)-gamma-aminopropylmethyldimethoxysilane (AEAPMDMS). DNA was loaded in 50 mM tris(hydroxylmethyl)aminomethane-EDTA buffer at pH 7.0 and eluted with 300 mM potassium phosphate solution at pH 10.0. Under optimal condition, a 6.0-cm monolithic column provided a capacity of 56 ng DNA with an extraction efficiency of 71 +/- 5.2% (X +/- RSD). When the amino silica monolithic column was applied to extract genomic DNA from the whole blood of crucian carp, an extraction efficiency of 52 +/- 5.6% (X +/- SD) was obtained by three extractions. Since the chaotropic-based sample loading and organic solvent wash steps were avoided in this procedure, the purified DNA was suitable for downstream processes such as PCR. This amino silica monolithic column was demonstrated to allow rapid and efficient DNA purification in microscale.
关键词[WOS]: BACTERIAL MAGNETIC PARTICLES ; SOLID-PHASE EXTRACTION ; NUCLEIC-ACIDS ; MICROFLUIDIC CHIPS ; PLASMID DNA ; PURIFICATION ; MICROCHIP ; SAMPLES ; SYSTEM ; FABRICATION
语种: 英语
WOS记录号: WOS:000269413400032
ISSN号: 1615-9306
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/7598
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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作者单位: 1.Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, Wuhan, Peoples R China

Recommended Citation:
Liu, Lijia; Yu, Shengbing; Yang, Shuixian; Zhou, Ping; Hu, Jiming; Zhang, Yibing.Extraction of genomic DNA using a new amino silica monolithic column,JOURNAL OF SEPARATION SCIENCE,2009,32(15-16):2752-2758
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