Determination of phosphorus impurity that directly affects quantification of microbial genomic DNA using inductively coupled plasma optical emission spectrometry.
We prepared genomic DNA from human placenta, Escherichia coli, and Bacillus subtilis using various DNA extraction methods and quantified the genomic DNA using ultraviolet (UV) spectrophotometry, capillary electrophoresis (CE), and inductively coupled plasma optical emission spectrometry (ICP-OES). Application of ICP-OES unexpectedly led to a serious overestimation of phosphorus in B. subtilis genomic DNA prepared using cetyltrimethyl ammonium bromide (CTAB). Further investigations using reversed-phase high-performance liquid chromatography (RP-HPLC), ultra-performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS), and (31)P nuclear magnetic resonance (NMR) identified the phosphorus impurity as lipoteichoic acid (LTA).
Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, Republic of Korea; Department of Bio-Analytical Science, University of Science & Technology, Daejeon 305-340, Republic of Korea.
Hyo-Jin
HJ
Yang
Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, Republic of Korea; Department of Bio-Analytical Science, University of Science & Technology, Daejeon 305-340, Republic of Korea.
Inchul
I
Yang
Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, Republic of Korea.
Jun-Hyuk
JH
Choi
Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, Republic of Korea.
Dukjin
D
Kang
Center for Inorganic Analysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, Republic of Korea.
Myung-Sub
MS
Han
Center for Bioanalysis, Division of Metrology for Quality of Life, Korea Research Institute of Standards and Science, Daejeon 305-340, Republic of Korea; Department of Bio-Analytical Science, University of Science & Technology, Daejeon 305-340, Republic of Korea. Electronic address: kimsk@kriss.re.kr.
Sook-Kyung
SK
Kim
D016477:Artifacts; D001412:Bacillus subtilis / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis*; D004340:Drug Contamination; D004926:Escherichia coli / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics*; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D008070:Lipopolysaccharides / Q000032:analysis; D013058:Mass Spectrometry; D010758:Phosphorus / Q000032:analysis*; D013054:Spectrophotometry, Atomic / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet; D013682:Teichoic Acids / Q000032:analysis
D004269:DNA, Bacterial; D008070:Lipopolysaccharides; D013682:Teichoic Acids; D010758:Phosphorus; C009900:lipoteichoic acid
Genomic DNA; Lipoteichoic acid; Phosphorus impurity