title: Use of fluorescent DNA-intercalating dyes in the analysis of DNA via ion-pair reversed-phase denaturing high-performance liquid chromatography.
abstract: SYBR Green 1 is an asymmetrical cyanine DNA-binding dye that provides an opportunity for increasing the sensitivity of nucleic acid detection when used in conjunction with gel electrophoresis. In this paper, we summarize the general properties and specific uses of SYBR green 1 in ion-pair reversed-phase denaturing high-performance liquid chromatography (IP DHPLC). We describe several applications for the WAVE DHPLC platform that illustrate the generic potential of such intercalating dyes in mutation detection and gene expression profiling. We show that SYBR Green 1 obviates the need to use end-labeled oligodeoxynucleotides for the sensitive detection of nucleic acids during chromatography. Moreover the incorporation of SYBR Green 1 into samples and elution buffers does not impair resolution and has no significant effect on the retention times of DNA fragments compared with dye-free DHPLC.
authors.affiliation: Department of Molecular Biology, The Transgenomic Research Laboratory, Krebs Institute, Firth Court, Western Bank, Sheffield S10 2TN, UK.
authors.forename: Ahmad R
authors.initials: AR
authors.lastname: Bahrami
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Dickman
authors.forename: Maryam M
authors.initials: MM
authors.lastname: Matin
authors.forename: John R
authors.initials: JR
authors.lastname: Ashby
authors.forename: Paul E
authors.initials: PE
authors.lastname: Brown
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Conroy
authors.forename: Gregory J S
authors.initials: GJ
authors.lastname: Fowler
authors.forename: James P
authors.initials: JP
authors.lastname: Rose
authors.forename: Qaiser I
authors.initials: QI
authors.lastname: Sheikh
authors.forename: Anthony T
authors.initials: AT
authors.lastname: Yeung
authors.forename: David P
authors.initials: DP
authors.lastname: Hornby
mesh_terms: D020137:Base Pair Mismatch; D002232:Carbocyanines / Q000737:chemistry; D002454:Cell Differentiation / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D053938:DNA Fragmentation; D018076:DNA, Complementary / Q000032:analysis; D005456:Fluorescent Dyes / Q000737:chemistry*; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D007364:Intercalating Agents / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: D002232:Carbocyanines; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D009838:Oligodeoxyribonucleotides; D004247:DNA