Capillary electrophoresis with dual laser detection in separation of amplified fragment length polymorphism fragments

Stefanowicz–Hajduk, Justyna; Filipowicz, Natalia; Kosiński, Igor; Ochocka, J. Renata

doi:10.1002/jssc.200900297

Cited by 8 publications

(2 citation statements)

References 17 publications

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“…The basic concept of the VP is that different DNA molecules have different optimum separation voltage conditions . Also, the DNA molecule can be detected with high sensitivity using an LIF detector .…”

Section: Introductionmentioning

confidence: 99%

Voltage‐programming‐based capillary gel electrophoresis for the fast detection of angiotensin‐converting enzyme insertion/deletion polymorphism with high sensitivity

Woo

Kim

Kang

2016

J of Separation Science

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A voltage-programming-based capillary gel electrophoresis method with a laser-induced fluorescence detector was developed for the fast and highly sensitive detection of DNA molecules related to angiotensin-converting enzyme insertion/deletion polymorphism, which has been reported to influence predisposition to various diseases such as cardiovascular disease, high blood pressure, myocardial infarction, and Alzheimer's disease. Various voltage programs were investigated for fast detection of specific DNA molecules of angiotensin-converting enzyme insertion/deletion polymorphism as a function of migration time and separation efficiency to establish the effect of voltage strength to resolution. Finally, the amplified products of the angiotensin-converting enzyme insertion/deletion polymorphism (190 and 490 bp DNA) were analyzed in 3.2 min without losing resolution under optimum voltage programming conditions, which were at least 75 times faster than conventional slab gel electrophoresis. In addition, the capillary gel electrophoresis method also successfully applied to the analysis of real human blood samples, although no polymorphism genes were detected by slab gel electrophoresis. Consequently, the developed voltage-programming capillary gel electrophoresis method with laser-induced fluorescence detection is an effective, rapid analysis technique for highly sensitive detection of disease-related specific DNA molecules.

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Section: Introductionmentioning

confidence: 99%

Voltage‐programming‐based capillary gel electrophoresis for the fast detection of angiotensin‐converting enzyme insertion/deletion polymorphism with high sensitivity

Woo

Kim

Kang

2016

J of Separation Science

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“…Because of the injection volume is quite small, it is essential to select a sensitive detector to improve detection sensitivity. Owing to high intensity, excellent selectivity and high monochromaticity of laser, LIF detection is considered as the best approach and reported most frequently in MCE because of the remarkable sensitivity [30,31]. In this work, we developed a microchip CE-LIF combined with onestep duplex RT-PCR method for rapid and specific detection of EV71 and CoxA16 in throat swab specimens.…”

Section: Introductionmentioning

confidence: 99%

Microchip capillary electrophoresis with laser‐induced fluorescence combined with one‐step duplex reverse‐transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens

Ruan

Sun

Chen

et al. 2015

J of Separation Science

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Enterovirus 71 and Coxsackievirus A16 are the main pathogens causing hand-foot-mouth disease. In this paper, microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse transcript-polymerase chain reaction has been developed for the detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens. The specific reverse transcription-polymerase chain reaction amplicons labeled with SYBR Orange were separated by microchip capillary electrophoresis and detected by laser induced fluorescence detector within 7 min. The intraday and interday relative standard deviation of migration time for DNA Marker was in the range of 1.36-2.94 and 2.78-3.96%, respectively. The detection limits were as low as 2.06 × 10(3) copies/mL for Enterovirus 71 and 5 × 10(3) copies/mL for Coxsackievirus A16. No cross-reactivity was observed with rotavirus, astrovirus, norovirus, and adenovirus, which showed good specificity of the method. This assay was validated using 100 throat swab specimens that were detected by real-time reverse-transcript polymerase chain reaction in parallel and the two methods produced the same results. This study provided a rapid, sensitive and specific method for the detection of Enterovirus 71 and Coxsackievirus A16, which make a contribution to significant time and cost saving for the identification and treatment of patients.

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Capillary Electrophoresis of DNA from Cannabis sativa for Correlation of Samples to Geographic Origin

Coyle

2011

Methods in Molecular Biology

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For routine genetic analysis of Cannabis sativa, two methods are currently in use, (a) AFLP; amplified fragment length polymorphism analysis and (b) STR; short tandem repeat analysis. The AFLP method used on capillary electrophoresis instrumentation is fully described in this chapter. AFLP analysis generates numerous nonspecific marker fragments for a complex DNA pattern and is available in kit format for quality assurance of reagents. This method is particularly useful when discerning the genetics of highly inbred plant species that may share much of the same DNA with only slight differences due to their common genetic background. AFLP analysis, however, is most effective on fresh or well-preserved plant specimens where the integrity of the DNA is high and the sample is a single source specimen (i.e., not a mixture of plants or different species).

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Capillary electrophoresis with dual laser detection in separation of amplified fragment length polymorphism fragments

Cited by 8 publications

References 17 publications

Voltage‐programming‐based capillary gel electrophoresis for the fast detection of angiotensin‐converting enzyme insertion/deletion polymorphism with high sensitivity

Voltage‐programming‐based capillary gel electrophoresis for the fast detection of angiotensin‐converting enzyme insertion/deletion polymorphism with high sensitivity

Microchip capillary electrophoresis with laser‐induced fluorescence combined with one‐step duplex reverse‐transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens

Capillary Electrophoresis of DNA from Cannabis sativa for Correlation of Samples to Geographic Origin

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