scholarly journals A comparison of DNA stains and staining methods for Agarose Gel Electrophoresis

2019 ◽  
Author(s):  
Andie C. Hall

ABSTRACTNucleic acid stains are necessary for Agarose Gel Electrophoresis (AGE). The commonly used but mutagenic Ethidium Bromide is being usurped by a range of safer but more expensive alternatives. These safe stains vary in cost, sensitivity and the impedance of DNA as it migrates through the gel. Modified protocols developed to reduce cost increase this variability. In this study, five Gel stains (GelRed™, GelGreen™, SYBR™ safe, SafeView and EZ-Vision®In-Gel Solution) two premixed loading dyes (SafeWhite, EZ-Vision®One) and four methods (pre-loading at 100x, pre-loading at 10x, precasting and post-staining) are evaluated for sensitivity and effect on DNA migration. GelRed™ was found to be the most sensitive while the EZ-Vision® dyes and SafeWhite had no discernible effect on DNA migration. Homemade loading dyes were as effective as readymade ones at less than 4% of the price. This method used less than 1% of the dye needed for the manufacturer recommended protocols. Thus, with careful consideration of stain and method, Gel stain expenditure can be reduced by over 99%.

Author(s):  
Qing Huang ◽  
Wei-Ling Fu

AbstractEthidium bromide (EB) is a mutagen and toxin that is widely used in the laboratory for visualization of nucleic acids. Safer nucleic acid stains, such as SYBR


2021 ◽  
Vol 8 ◽  
Author(s):  
Caiyun Huo ◽  
Donghai Li ◽  
Zhenguo Hu ◽  
Guiping Li ◽  
Yanxin Hu ◽  
...  

Avibacterium paragallinarum, the pathogen of infectious coryza, caused a highly contagious respiratory disease that poses a serious threat to chickens. Hence, it is necessary to do diagnostic screening for Av. paragallinarum. Existing technologies have been used for Av. paragallinarum testing, which, however, have some drawbacks such as time consuming and expensive that require well-trained personnel and sophisticated infrastructure, especially when they are limitedly feasible in some places for lack of resources. Nucleic acid hybridization-based lateral flow assay (LFA) is capable of dealing with these drawbacks, which is attributed to the advantages, such low cost, rapid, and simple. However, nucleic acid determination of Av. paragallinarum through LFA method has not been reported so far. In this study, we developed a novel LFA method that employed gold nanoparticle probes to detect amplified Av. paragallinarum dsDNA. Compared with agarose gel electrophoresis, this LFA strip was inexpensive, simple- to- use, and time- saving, which displayed the visual results within 5–8 min. This LFA strip had higher sensitivity that achieved the detection limit of 101 CFU/ml compared with 102 CFU/ml in agarose gel electrophoresis. Besides, great sensitivity was also shown in the LFA strip, and no cross reaction existed for other bacteria. Furthermore, Av. paragallinarum in clinical chickens with infectious coryza were perfectly detected by our established LFA strip. Our study is the first to develop the LFA integrated with amplification and sample preparation techniques for better nucleic acid detection of Av. paragallinarum, which holds great potential for rapid, accurate, and on-site determination methods for early diagnosis of Av. paragallinarum to control further spreading.


1998 ◽  
Vol 8 (4) ◽  
pp. 227-233 ◽  
Author(s):  
T. N. Ferraro ◽  
J. F. Schill ◽  
C. Ballas ◽  
N. Mulholland ◽  
G. T. Golden ◽  
...  

2009 ◽  
Vol 71-73 ◽  
pp. 159-162 ◽  
Author(s):  
Carla M. Zammit ◽  
L.A. Mutch ◽  
Helen R. Watling ◽  
Elizabeth L.J. Watkin

Various methods of nucleic acid (NA) extraction were investigated with the aim of developing a quantitative method of NA extraction from five representative strains of biomining microorganisms. The process of removing cells from mineral surfaces, lysing microorganisms, precipitating NA and purifying RNA were analysed. The success of each method was examined spectrophotometrically, by agarose gel electrophoresis and PCR or quantitative real time PCR (qPCR). The most important step was shown to be cellular lysis, which principally impacted on the quantity of NA extracted from each strain. The quantity and quality of extracted NA was highly dependent on the method of NA precipitation. This study resulted in the development of a NA extraction method that reliably and reproducibly extracted NA from five strains of biomining microorganisms.


2006 ◽  
Vol 17 (16) ◽  
pp. 4263-4269 ◽  
Author(s):  
Alexandre A Vetcher ◽  
Srimeenakshi Srinivasan ◽  
Ivan A Vetcher ◽  
Semen M Abramov ◽  
Mikhail Kozlov ◽  
...  

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