Light‐emitting diode induced fluorescence (LED‐IF) detection design for a pen‐shaped cartridge based single capillary electrophoresis system

Abstract

CGE is a well-established separation technique for the analysis of biologically important molecules such as nucleic acids. The inherent high resolving power, rapid analysis times, excellent detection sensitivity, and quantification capabilities makes this method favorable compared to conventional manual polyacrylamide and agarose slab gel electrophoresis techniques. In this paper we introduce a novel single-channel capillary gel electrophoresis system with LED-induced fluorescence detection also utilizing a compact pen-shaped capillary cartridge design for automatic analysis of samples from a 96-well plate. To evaluate the suitability of the system, 1000 genomic DNA(gDNA) samples were analyzed in gel filled capillaries and detected by the microball ended excitation and emission optical fiber based LED-induced fluorescence detection system. Excellent migration time reproducibility of RSD <0.75% was obtained over the course of 1000 runs. The system rapidly distinguished between intact and degraded gDNA samples, therefore provided important information if they could be used for downstream quantitative PCR processing where high-quality intact gDNA was key. We envision that this novel system design will rapidly find new applications in both research and clinical diagnostic laboratories as a highly sensitive and easy to use bio-analytical approach.

Calcium mediated DNA binding in non-lamellar structures formed by DOPG/glycerol monooleate

“…The length of DNA fragments was determined by micro-capillary electrophoresis using single-channel Qsep 100 Bio-Fragment Analyzer (BiOptic Inc., Taiwan). A disposable pen-shaped cartridge was inserted and other running buffers were loaded according to the manufacturer's instructions (Kerékgyártó et al 2012). DNA was diluted to concentration of 89.90 ng.L -1 for analysis.…”

Combining Direct PCR Technology and Capillary Electrophoresis for an Easy-to-Operate and Highly Sensitive Infectious Disease Detection System for Shrimp

“…CE has been applied to various high-throughput nucleic acid analysis applications, including genotyping and pathogen detection, due to its high sensitivity, superior accuracy, speed, and automation [23][24][25]. The recent development of lightweight and portable CE machines makes this powerful tool an excellent apparatus for on-site nucleic acid detection [26]. However, regardless of the detection method, a nucleic acid purification step is required before PCR, which is inconvenient for on-site operation and can easily result in sample contamination during pathogen detection in aquaculture.…”

Design and evaluation of capillary coupled with optical fiber light‐emitting diode induced fluorescence detection for capillary electrophoresis

“…So far, CE has been widely used in DNA sequencing , protein analysis , saccharides analysis , pharmaceutical analysis , food analysis , chiral separation , and so on. A variety of detectors have been developed to improve the sensitivity, such as UV absorption , chemiluminescence , electrochemical , MS , LIF , and LED‐induced fluorescence (LED‐IF) . Though it is accepted that LIF is one of the most sensitive detection methods, the detector uses laser as excitation light source, which has certain shortcomings as the following: high cost, big volume, and limited suitable wavelengths to excite the analytes.…”

“…The proposed system was successfully applied to separate and determine riboflavin in real sample.Abbreviations: CCOF-LED-IFD or CCOF, capillary coupled with optical fiber LED-induced fluorescence detector; IC-COF-LED-IFD or COF, in-capillary common optical fiber LEDinduced fluorescence detector; LED-IF, LED-induced fluorescence; RF, riboflavin So far, CE has been widely used in DNA sequencing [3,4], protein analysis [5,6], saccharides analysis [7,8], pharmaceutical analysis [9, 10], food analysis [11,12], chiral separation [13], and so on. A variety of detectors have been developed to improve the sensitivity, such as UV absorption [14][15][16], chemiluminescence [17], electrochemical [18], MS [19,20], LIF [21][22][23], and LED-induced fluorescence (LED-IF) [24,25]. Though it is accepted that LIF is one of the most sensitive detection methods, the detector uses laser as excitation light source, which has certain shortcomings as the following: high cost, big volume, and limited suitable wavelengths to excite the analytes.…”