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  • 1
    Electronic Resource
    Electronic Resource
    A three-wavelength labeling approach for DNA sequencing using energy transfer primers and capillary electrophoresis (1998)
    Weinheim : Wiley-Blackwell
    Electrophoresis 19 (1998), S. 1403-1414 
    Details
    ISSN: 0173-0835
    Keywords: DNA sequencing ; Capillary electrophoresis ; Energy transfer primers ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Capillary electrophoresis DNA sequencing has been accomplished by using four different energy transfer primers and three fluorescence detection channels. Methods have also been developed to deconvolve the three-color data into the four base concentrations. The nonnegative least squares and model selection method resulted in the best accuracy. The three-color data were compared to sequencing data obtained using four detection channels and four energy transfer primers. The average accuracy rates obtained over three 500 base M13mp18 runs using three-color coding were 96% including 18 uncallable compressions and 99.6% if these compressions are excluded. The average accuracy rate obtained using four-color coding was 96.3% including 18 uncallable compressions and 99.9% if these compressions are excluded. Although it is unlikely that three-color schemes will replace four-color sequencing, these methods have exposed basic concepts that will be useful in the development of higher-order multiplex coding methods for DNA analysis.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150190835
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    DNA sequencing using a four-color confocal fluorescence capillary array scanner (1996)
    Weinheim : Wiley-Blackwell
    Electrophoresis 17 (1996), S. 1852-1859 
    Details
    ISSN: 0173-0835
    Keywords: DNA sequencing ; Capillary electrophoresis ; Confocal fluorescence detection ; Energy transfer primers ; Mitochondrial DNA polymorphisms ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The design, construction and operation of a four-color capillary array electrophoresis scanner are presented. The use of sensitive energy transfer primers facilitates four-color detection of the DNA sequencing fragments following excitation at a single laser wavelength (488 nm). This scanner collects fluorescence data from up to 25 capillaries in parallel. The resulting four-color image files are automatically reduced to four-color line plots, and a base-calling program (Sax) is used to call the sequence. The performance of this system for DNA sequencing is demonstrated by examining twelve different motifs of the hypervariable region I of human mitochondrial (mt) DNA obtained from a Sierra Leone population.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150171209
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  • 3
    Electronic Resource
    Electronic Resource
    Femtosecond time-resolved spectroscopy of the primary photochemistry of phytochrome (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Biospectroscopy 3 (1997), S. 421-433 
    Details
    ISSN: 1075-4261
    Keywords: plant photoreceptor ; photoisomerization mechanism ; excited state lifetime ; fluorescence quantum yield ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: Time-resolved absorption spectra of Pr phytochrome were obtained using a regeneratively amplified femtosecond titanium : sapphire laser system. The early time transient absorption spectra are comprised of prompt Pr photobleaching, stimulated emission, and excited-state absorption features that decay with a 24 ps time constant that matches the ground state appearance time of the primary photoproduct. Based on the 5 ns radiative lifetime calculated from the absorption and spontaneous emission spectra and the fluorescence quantum yield of 5.5 (± 0.5) × 10-3, we calculate an excited-state lifetime of 28 ps that agrees well with the directly determined lifetime. The transient absorption spectra are consistent with a primary photochemical reaction quantum yield of 0.15, and the absorption spectrum of the primary photoproduct closely resembles that of the low-temperature trapped intermediate, lumi-R. We conclude that the primary photoisomerization, which is believed to be a Z,syn → E,syn isomerization of the C15=C16 chromophore bond, occurs in 24 ps. © 1997 John Wiley & Sons, Inc. Biospectroscopy 3: 421-433, 1997
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Microsatellite-based cancer detection using capillary array electrophoresis and energy-transfer fluorescent primers (1997)
    Weinheim : Wiley-Blackwell
    Electrophoresis 18 (1997), S. 1742-1749 
    Details
    ISSN: 0173-0835
    Keywords: Cancer diagnosis ; Short tandem repeat ; Capillary electrophoresis ; Energy-transfer primers ; Microsatellites ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The development of sensitive, rapid, and accurate methods and apparatus for high-throughput short tandem repeat (STR) analysis will be critical for the use of microsatellite alteration in cancer screening. Here we show that STR-based bladder cancer diagnosis can be performed using capillary array electrophoresis and two-color labeling with energy-transfer (ET) fluorescent primers. Rapid (≤35 min) separations are achieved on capillary arrays using replaceable separation matrices and the allelic ratios are quantitatively determined with a precision of ± 10%. With this precision, a variation of 20% was considered diagnostically significant. These methods provide a significant improvement in the speed, ease, and precision of STR analyses compared to slab gel electrophoresis.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150181007
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  • 5
    Electronic Resource
    Electronic Resource
    High-resolution capillary array electrophoretic sizing of multiplexed short tandem repeat loci using energy-transfer fluorescent primers (1996)
    Weinheim : Wiley-Blackwell
    Electrophoresis 17 (1996), S. 1485-1490 
    Details
    ISSN: 0173-0835
    Keywords: Capillary array electrophoresis ; Short tandem repeat ; Energy transfer primers ; Microsatellites ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Short tandem repeat regions (STRs) from the polymorphic loci VWFA, THO1, TPO and CSF were amplified by the multiplex polymerase chain reaction (PCR) and analyzed by capillary array electrophoresis with fluorescence detection of energy transfer (ET) labels. The fluorescent ET primers are labeled with one fluorescein at the 5′ end and a second fluorescein at the position of the 7th or 9th (modified) base to produce fragments that fluoresce in the green (λmax = 525 nm). M13 A-track sequencing fragments, used as an internal sizing standard, were generated with a universal primer that has a donor fluorescein at the 5′ end and a rhodamine acceptor at the position of the 11th (modified) base to produce fragments fluorescing in the red (〉 590 nm). The labeled DNA fragments were excited at 488 nm, and the fluorescence was detected with a two-color confocal fluorescence scanner. Separations were performed on arrays of hollow fused silica capillaries filled with denaturing and replaceable hydroxyethyl cellulose sieving matrices. Separations were complete in less than 50 min, and single base resolution as well as reproducible STR sizing was achieved. The relative standard deviation in sizing was below 0.6%. This work establishes the feasibility of high-resolution, high-speed and high-throughput STR typing of single-stranded DNA fragments using capillary array electrophoresis.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150170913
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  • 6
    Electronic Resource
    Electronic Resource
    Raman spectroscopy with intensified vidicon detectors: A study of intact bovine lens proteins (1978)
    Chichester [u.a.] : Wiley-Blackwell
    Journal of Raman Spectroscopy 7 (1978), S. 349-352 
    Details
    ISSN: 0377-0486
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Physics
    Notes: Raman spectra of intact ocular lenses contain important structural information related to aging and cataract formation. However, the direct application of Raman spectroscopy to human eyes for cataract diagnosis has been considered unsafe because of the high laser power (∼100 mW) required to generate Raman spectra using the photomultiplier-photon counting (PMT-PC) detection method. The clinical application of Raman spectroscopy has become more likely with the development of sensitive multichannel optical detectors such as the ‘silicon intensified target’ (SIT) coupled to an optical multichannel analyzer (OMA). We have incorporated the SIT in a novel mismatched subtractive dispersion double monochromator that has optimum stray light (〈5×10-9), throughput, and resolution for these experiments. It is demonstrated that the new SIT-OMA detection method is capable of obtaining high-quality Raman spectra of an intact bovine lens with a laser power (∼1 mW) significantly below the threshold for retinal damage. The multichannel and photon counting detection systems have been carefully compared under high and low light level conditions so that the advantages of the SIT-OMA system for other Raman experiments may be easily evaluated.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jrs.1250070613
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  • 7
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    Integration of a Micro-Chip Amino Acid Chirality Detector into the MOD Instrument Concept (2004)
    In:  CASI
    Details
    Publication Date: 2019-07-13
    Description: The MOD (Mars Organic Detector) instrument concept consists of a sublimation apparatus for organic compound isolation connected to a microfabricated microfluidic analyzer containing a sipper, pumps and a separation channel for organic compound characterization. The target organic compounds are amino acids and polycyclic aromatic hydrocarbons (PAHs). Solid samples are placed within the sublimation apparatus and heated to release organic compounds which sublime onto a cold finger. Half of the cold finger is coated with fluorescamine. which reacts with amino acids and other primary amines to generate an intense fluorescent derivative while the other half is uncoated and is used to directly detect PAH fluorescence, A capillary sipper is then used to dissolve and sample the labeled amino acids and integrated microfabricated pumps transport the labeled amino acids to the chip for analysis. The sample is separated using capillary zone electrophoresis (CZE) together with chiral dextrins to determine amino acid composition and chirality. During the grant period, the following steps have been completed toward the development of a robust instrument and chemistry.
    Keywords: Electronics and Electrical Engineering
    Format: application/pdf
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