Premium
Extraction of genomic DNA using a new amino silica monolithic column
Author(s) -
Liu Lijia,
Yu Shengbing,
Yang Shuixian,
Zhou Ping,
Hu Jiming,
Zhang Yibing
Publication year - 2009
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200900208
Subject(s) - monolithic hplc column , chromatography , elution , chaotropic agent , chemistry , extraction (chemistry) , tris , dna , microscale chemistry , polymerization , dna extraction , high performance liquid chromatography , polymer , biochemistry , polymerase chain reaction , organic chemistry , mathematics education , mathematics , gene
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 ‐(β‐aminoethyl)‐γ‐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 ± RSD) 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.