In vitro selection of DNA to chloroaromatics using magnetic microbead-based affinity separation and fluorescence detection.
In vitro selection (SELEX) of DNA ligands to the chloroaromatics, 4-chloroaniline (4-CA), 2,4,6-trichloroaniline (TCA) and pentachlorophenol (PCP), was performed by a novel method utilizing magnetic beads (MBs) having a linker arm for immobilization. Use of MBs was advantageous in obviating elution and precipitation of DNA as conjugated MBs with surface-captured template DNA could be directly added to PCR mixtures. In addition, a simplified PCR scheme requiring only one type of primer and a rapid fluorescence microscopic method for assessing nucleic acid binding after each round of SELEX were demonstrated.
Bruno
J G
JG
Applied Research Associates, Tyndall Air Force Base, Florida 32403, USA.
D000814:Aniline Compounds / Q000378:metabolism*; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism*; D017931:DNA Primers; D006843:Hydrocarbons, Chlorinated / Q000378:metabolism*; D008024:Ligands; D008280:Magnetics; D008856:Microscopy, Fluorescence; D008863:Microspheres; D009838:Oligodeoxyribonucleotides / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction
D000814:Aniline Compounds; D017931:DNA Primers; D006843:Hydrocarbons, Chlorinated; D008024:Ligands; D009838:Oligodeoxyribonucleotides; D004247:DNA; C045381:2,4,6-trichloroaniline; C027673:2,3,4,5,6-pentachloroaniline; C004658:4-chloroaniline