Preparative isolation of polymerase chain reaction products using mixed-mode chromatography.
The polymerase chain reaction (PCR) has become one of the most useful techniques in molecular biology laboratories around the world. The purification of the target DNA product is often challenging, however, and most users are restricted to employing available commercial kits. The recent developments in mixed-mode chromatography have shown higher selectivity for a variety of nucleic acid-containing samples. Capto Adhere is a mixed-mode chromatography resin that offers a high-selectivity ligand and is here applied for the purification of amplified DNAs from PCR mixtures in a 10-min single step, with yields above 95%, high linearity, and high precision for different concentrations.
Department of Pure and Applied Biochemistry, Lund University, SE-221 00 Lund, Sweden; Health Sciences Research Centre, University of Beira Interior, 6200 506 Covilhã, Portugal. Electronic address: tiago.matos@tbiokem.lth.se.
T
T
Matos
Department of Pure and Applied Biochemistry, Lund University, SE-221 00 Lund, Sweden; Health Sciences Research Centre, University of Beira Interior, 6200 506 Covilhã, Portugal.
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G
Silva
Health Sciences Research Centre, University of Beira Interior, 6200 506 Covilhã, Portugal.
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JA
Queiroz
Department of Pure and Applied Biochemistry, Lund University, SE-221 00 Lund, Sweden.
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L
Bülow
D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D018744:DNA, Plant / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004274:DNA, Recombinant / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D018744:DNA, Plant; D004274:DNA, Recombinant; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D004247:DNA
Double-stranded DNA; Hydrophobic interaction chromatography; Ion-exchange chromatography; Plasmid DNA; Polymerase chain reaction; Single-stranded DNA