title: Temperature-independent Detection of Heteroduplex and Homoduplex Fragments Applying Poly(glycidylmethacryate-co-divinylbenzene-based) Monoliths Modified to Strong Anion-exchanger.
abstract: Monoliths of poly(glycidylmethacrylate-co-divinylbenzene) were prepared in the confines of presilanized borosilicate glass columns (100 × 3 mm I.D.). These monoliths were surface modified into strong anion-exchangers with hydrochloric acid (10%) and triethylamine, successively. The strong anion-exchanger established good separation of 5-phosphorylated oligodeoxythymidylic acids fragments [d(pT)(12-18)]. Moreover, heteroduplex and homoduplex fragments of a low-range mutation standard [of STS marker from the Y-chromosome (209 bp)] were separated at ambient and elevated temperatures using sodium phosphate buffer and a gradient former of sodium chloride, in anion-exchange high-performance liquid chromatography (AE-HPLC). This is a step forward for mutation detection as temperature-independent method, which is not the case in denatured ion-paired reversed-phase chromatography (D-IP-RP-HPLC), where mutation detection is temperature critical and might be bypassed if temperature changes slightly. Finally, reproducibility check from run-to-run and monolith-to-monolith showed a relative standard deviation (RSD) of less than 2%.
authors.forename: Said H
authors.initials: SH
authors.lastname: Lubbad
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D010539:Permeability; D010766:Phosphorylation; D011109:Polymethacrylic Acids / Q000737:chemistry*; D011145:Polyvinyls / Q000737:chemistry*; D016062:Porosity; D015203:Reproducibility of Results; D017640:Silicates / Q000737:chemistry; D013499:Surface Properties; D013696:Temperature
chemical_list: D009838:Oligodeoxyribonucleotides; D011109:Polymethacrylic Acids; D011145:Polyvinyls; D017640:Silicates; C027903:oligo (dT); C516630:poly(glycidyl methacrylate-co-divinylbenzene); D004247:DNA