Rapid sizing of short tandem repeat alleles using capillary array electrophoresis and energy-transfer fluorescent primers

Analytical Chemistry
Y WangR A Mathies

Abstract

Genetic typing of the short tandem repeat (STR) polymorphism HUMTHO1 has been performed using capillary array electrophoresis and energy-transfer fluorescent dye-labeled polymerase chain reaction primers. Target alleles were amplified by use of primers labeled with one fluorescein at the 5' end and another fluorescein at the position of the 15th (modified) base to produce fragments that fluoresce in the green (lambda max = 525 nm). Unknown alleles were electrophoretically separated together with a standard ladder made up of alleles having 6, 7, 8, and 9 four-base pair repeats, each of which was amplified with an energy-transfer primer having a donor fluorescein at the 5' end and a rhodamine acceptor at the position of the 7th (modified) base to produce standard fragments fluorescing in the red (> 590 nm). Separations were performed on arrays of hollow fused-silica capillaries filled with a replaceable sieving matrix consisting of 0.8% hydroxyethyl cellulose plus 1 microM 9-aminoacridine to enhance the resolution. The labeled DNA fragments were excited at 488 nm, and the fluorescence was detected with a two-color confocal fluorescence scanner. Separations are complete in less than 20 min and allow sizing with an average absolute...Continue Reading

Citations

Feb 10, 1998·Electrophoresis·W M Sunada, H W Blanch
Mar 25, 1998·Electrophoresis·J W Mitchell, D Walsh
Aug 9, 1996·Journal of Chromatography. B, Biomedical Applications·P BelgraderM A Marino
Dec 2, 2000·Journal of Chromatography. a·A P SassiH H Hooper
Nov 22, 1997·Trends in Biotechnology·K R MitchelsonL J Kricka
Jun 6, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·X Gong, E S Yeung
Sep 24, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·N Zhang, E S Yeung
Oct 27, 2001·Biosensors & Bioelectronics·H NagaiE Tamiya
Feb 1, 1997·Current Opinion in Biotechnology·A N Glazer, R A Mathies
May 9, 1995·Proceedings of the National Academy of Sciences of the United States of America·J JuR A Mathies
Jun 15, 2007·Pharmacogenomics·Gudrun H ReedCarl T Wittwer
Sep 18, 2012·Biosensors & Bioelectronics·Hui ZhouRu-Qin Yu
Mar 8, 2005·Analytical and Bioanalytical Chemistry·Jianhua QinBingcheng Lin
Feb 12, 2010·The World Journal of Biological Psychiatry : the Official Journal of the World Federation of Societies of Biological Psychiatry·André TadićNorbert Dahmen
Feb 21, 1998·Genome Research·H WenzJ W Efcavitch
Mar 24, 2009·BioTechniques·Christopher D IntemannChristian G Meyer
Nov 2, 2007·Chemical Reviews·Karel Kleparník, Petr Bocek

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