Ultra-high-speed DNA sequencing using capillary electrophoresis chips.
AuthorsWoolley AT, Mathies RA
JournalAnal Chem
PubMed ID8644919
'DNA sequencing has been performed on microfabricated capillary electrophoresis chips. DNA separations were achieved in 50 x 8 microns cross-section channels microfabricated in a 2 in. x 3 in. glass sandwich structure using a denaturing 9% T, 0% C polyacrylamide sieving medium. DNA sequencing fragment ladders were produced and fluorescently ... More
Real-time PCR in the microbiology laboratory.
AuthorsMackay IM
JournalClin Microbiol Infect
PubMed ID15008940
'Use of PCR in the field of molecular diagnostics has increased to the point where it is now accepted as the standard method for detecting nucleic acids from a number of sample and microbial types. However, conventional PCR was already an essential tool in the research laboratory. Real-time PCR has ... More
Energy-transfer fluorescent reagents for DNA analyses.
AuthorsGlazer AN, Mathies RA
JournalCurr Opin Biotechnol
PubMed ID9013654
'Fluorescence resonance energy transfer has facilitated the development of a new class of high-performance fluorescent labeling reagents for multiplex analyses of nucleic acids. The enhanced emission of energy transfer (ET) primers has provided a decadic improvement in the performance of automated DNA sequencers. The emission spectral purity of ET primers ... More
Energy transfer primers with 5- or 6-carboxyrhodamine-6G as acceptor chromophores.
AuthorsHung SC, Ju J, Mathies RA, Glazer AN
JournalAnal Biochem
PubMed ID8660606
'Energy-transfer (ET) fluorescent primers for DNA sequencing and multiplex genetic analysis (Ju, J., Ruan, C., Fuller, C. W., Glazer, A. N., and Mathies, R. A. (1995) Proc. Natl. Acad. Sci. USA 92, 4347-4351) are named according to the convention D-N-A, where D is the donor, N is the number of ... More
Fluorescence excitation transfer immunoassay (FETI).
AuthorsUllman EF, Khanna PL
JournalMethods Enzymol
PubMed ID7033726
Detection of specific DNA sequences by fluorescence amplification: a color complementation assay.
AuthorsChehab FF, Kan YW
JournalProc Natl Acad Sci U S A
PubMed ID2594760
We have developed a color complementation assay that allows rapid screening of specific genomic DNA sequences. It is based on the simultaneous amplification of two or more DNA segments with fluorescent oligonucleotide primers such that the generation of a color, or combination of colors, can be visualized and used for ... More
Simultaneous detection of multiple point mutations using fluorescence-coupled competitive primer extension.
AuthorsFauser S, Wissinger B
JournalBiotechniques
PubMed ID9149883
We report the development of a method for the simultaneous genotyping of several distinct nucleotide positions by means of fluorescence-coupled competitive primer extension. We demonstrate the application of this method for the simultaneous detection of three point mutations in the human mitochondrial genome, at nucleotide positions 3460, 11778 and 14484, ... More
DNA sequencing by capillary electrophoresis: use of a two-laser-two-window intensified diode array detection system.
This paper presents the principles of an instrument designed for DNA sequencing using the standard four-dye-labeled primer approach. The method is based on capillary electrophoresis with laser-induced fluorescence and an intensified diode array detector. An important goal of the instrument design has been a detection system that possesses high sensitivity ... More
Effect of primary and secondary structure of oligodeoxyribonucleotides on the fluorescent properties of conjugated dyes.
We studied fluorescence intensity, polarization and lifetime of some commonly used fluorophores conjugated to oligodeoxyribonucleotides with different primary and secondary structures. We found that fluorescence intensity can increase or decrease upon hybridization of the labeled strand to its complement depending on the sequence and position of the fluorophore. Up to ... More
Influence of fluorophor dye labels on the migration behavior of polymerase chain reaction--amplified short tandem repeats during denaturing capillary electrophoresis.
AuthorsHahn M, Wilhelm J, Pingoud A
JournalElectrophoresis
PubMed ID11545394
The determination of the length of polymerase chain reaction (PCR)-amplified short tandem repeats (STRs) by denaturing capillary electrophoresis (CE) is a standard procedure for purposes of genotyping. We show that dye-specific mobility anomalies exist for 5'-fluorophor-labelled single-stranded DNA (ssDNA) fragments in CE using the performance-optimized polymer 4 (POP4) buffer sieving ... More
High-resolution liquid chromatography of fluorescent dye-labeled nucleic acids.
Using 100 mM of triethylammonium acetate as ion-pairing reagent, phosphodiester oligonucleotides labeled fluorescently at their 5' terminus could be separated successfully on alkylated nonporous 2.3-microns poly(styrene-divinylbenzene) particles by means of high-resolution liquid chromatography. Applying excitation wavelengths of 490, 520, 550, and 575 nm, respectively, optimum sensitivity was achieved for the ... More
Optimization of spectroscopic and electrophoretic properties of energy transfer primers.
AuthorsHung SC, Mathies RA, Glazer AN
JournalAnal Biochem
PubMed ID9324944
We have synthesized and characterized the spectroscopic properties of 56 energy transfer (ET) fluorescent dye-labeled primers differing in (i) the spacing between the donor and acceptor, (ii) the nature of the spacer (either oligonucleotide or polydideoxyribose phosphate), (iii) the primer sequence (M13 (-40), M13 (-21), M13 reverse, SP6, T3, and ... More
Design and synthesis of fluorescence energy transfer dye-labeled primers and their application for DNA sequencing and analysis.
AuthorsJu J, Kheterpal I, Scherer JR, Ruan C, Fuller CW, Glazer AN, Mathies RA
JournalAnal Biochem
PubMed ID8678292
We have designed and synthesized fluorescent oligonucleotide primers having improved fluorescence and electrophoretic properties by exploiting the concept of resonance fluorescence energy transfer (ET). These primers carry a fluorescein derivative at the 5' end as a common fluorescence donor and other fluorescein and rhodamine derivatives attached to a modified thymidine ... More
Stains, labels and detection strategies for nucleic acids assays.
AuthorsKricka LJ
JournalAnn Clin Biochem
PubMed ID11928759
Selected developments and trends in stains, labels and strategies for detecting and measuring nucleic acids (DNA, RNA) and related molecules [e.g. oligo(deoxy)nucleotides, nucleic acid fragments and polymerase chain reaction products] are surveyed based on the literature in the final decade of the 20th century (1991-2000). During this period, important families ... More
Differential gene regulation in human versus rodent hepatocytes by peroxisome proliferator-activated receptor (PPAR) alpha. PPAR alpha fails to induce peroxisome proliferation-associated genes in human cells independently of the level of receptor expresson.
AuthorsLawrence JW, Li Y, Chen S, DeLuca JG, Berger JP, Umbenhauer DR, Moller DE, Zhou G
JournalJ Biol Chem
PubMed ID11418601
We compared the ability of rat and human hepatocytes to respond to fenofibric acid and a novel potent phenylacetic acid peroxisome proliferator-activated receptor (PPAR) alpha agonist (compound 1). Fatty acyl-CoA oxidase (FACO) activity and mRNA were increased after treatment with either fenofibric acid or compound 1 in rat hepatocytes. In ... More