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Extraction of genomic DNA using a new amino silica monolithic column
Liu, Lijia1; Yu, Shengbing1; Yang, Shuixian1; Zhou, Ping1; Hu, Jiming1; Zhang, Yibing2; Zhou, P, Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China
2009-08-01
Source PublicationJOURNAL OF SEPARATION SCIENCE
ISSN1615-9306
Volume32Issue:15-16Pages:2752-2758
AbstractA 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.; 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.
SubtypeArticle
KeywordDna Purification Genomic Dna Monolithic Column Solid-phase Extraction
Department[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
Subject AreaChemistry ; Analytical
DOI10.1002/jssc.200900208
WOS HeadingsScience & Technology ; Physical Sciences
Funding OrganizationNatural Science Foundation of China [20875073] ; Natural Science Foundation of China [20875073] ; Natural Science Foundation of China [20875073] ; Natural Science Foundation of China [20875073]
Indexed BySCI
Language英语
WOS Research AreaChemistry
WOS SubjectChemistry, Analytical
WOS IDWOS:000269413400032
WOS KeywordBACTERIAL MAGNETIC PARTICLES ; SOLID-PHASE EXTRACTION ; NUCLEIC-ACIDS ; MICROFLUIDIC CHIPS ; PLASMID DNA ; PURIFICATION ; MICROCHIP ; SAMPLES ; SYSTEM ; FABRICATION
Funding OrganizationNatural Science Foundation of China [20875073] ; Natural Science Foundation of China [20875073] ; Natural Science Foundation of China [20875073] ; Natural Science Foundation of China [20875073]
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Cited Times:12[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/7598
Collection期刊论文
Corresponding AuthorZhou, P, Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China
Affiliation1.Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, Wuhan, Peoples R China
Recommended Citation
GB/T 7714
Liu, Lijia,Yu, Shengbing,Yang, Shuixian,et al. Extraction of genomic DNA using a new amino silica monolithic column[J]. JOURNAL OF SEPARATION SCIENCE,2009,32(15-16):2752-2758.
APA Liu, Lijia.,Yu, Shengbing.,Yang, Shuixian.,Zhou, Ping.,Hu, Jiming.,...&Zhou, P, Wuhan Univ, Coll Chem & Mol Sci, Wuhan 430072, Peoples R China.(2009).Extraction of genomic DNA using a new amino silica monolithic column.JOURNAL OF SEPARATION SCIENCE,32(15-16),2752-2758.
MLA Liu, Lijia,et al."Extraction of genomic DNA using a new amino silica monolithic column".JOURNAL OF SEPARATION SCIENCE 32.15-16(2009):2752-2758.
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