scholarly journals Principles of Nucleic Acid Separation by Agarose Gel Electrophoresis

10.5772/38654 ◽  
2012 ◽  
Author(s):  
Muhittin Ylmaz ◽  
Cem Ozic ◽  
lhami Gok
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis

Comparative analysis of the DNA staining efficiencies of different fluorescent dyes in preparative agarose gel electrophoresis

2005 ◽  
Vol 43 (8) ◽  
Author(s):  
Qing Huang ◽  
Wei-Ling Fu
Keyword(s):  
Comparative Analysis ◽  
Nucleic Acid ◽  
Nucleic Acids ◽  
Ethidium Bromide ◽  
Gel Electrophoresis ◽  
Fluorescent Dyes ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Dna Staining

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

scholarly journals A Novel Lateral Flow Assay for Rapid and Sensitive Nucleic Acid Detection of Avibacterium paragallinarum

2021 ◽  
Vol 8 ◽  
Author(s):  
Caiyun Huo ◽  
Donghai Li ◽  
Zhenguo Hu ◽  
Guiping Li ◽  
Yanxin Hu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nucleic Acid Hybridization ◽  
Nucleic Acid Detection ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Infectious Coryza ◽  
Avibacterium Paragallinarum

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.

Nucleic Aacid Extraction from Biomining Microorganisms

2009 ◽  
Vol 71-73 ◽  
pp. 159-162 ◽  
Author(s):  
Carla M. Zammit ◽  
L.A. Mutch ◽  
Helen R. Watling ◽  
Elizabeth L.J. Watkin
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Extraction Method ◽  
Quantitative Method ◽  
Agarose Gel ◽  
Mineral Surfaces ◽  
Agarose Gel Electrophoresis ◽  
Quantitative Real Time Pcr ◽  
Cellular Lysis

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.

Fractionation of SWNT/nucleic acid complexes by agarose gel electrophoresis

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

scholarly journals Enhancement of Recombinase-Aided Amplification (Raa) Assay by Betaine and Pullulan

2021 ◽  
Author(s):  
Jinrong Wang ◽  
Guowei Song ◽  
Yue Ming ◽  
Jing Pan ◽  
Ruiqing Zhang ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Nucleic Acid Amplification ◽  
Basic Method ◽  
Clinical Samples ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Fluorescence Methods ◽  
Threshold Time ◽  
First Time

Abstract In this research, nucleic acid amplification enhancers suitable for recombinase-aided amplification (RAA) assay were studied for the first time, and amplification of a long-fragment (509 bp) by the RAA assay was initially explored. Using recombinant plasmids and clinical samples, RAA fluorescence and basic methods were used to evaluate the efficacy. The fluorescence method was evaluated by threshold time (TT) and fluorescence value, and the basic method was interpreted by 2% agarose gel electrophoresis. Taking a previously established RAA assay for HPV18 as an example, we demonstrated that the addition of 0.2, 0.4, and 0.6 M betaine and 10% pullulan could enhance RAA. The new RAA assays with betaine and pullulan were called B-RAA and P-RAA, respectively. In the B-RAA and P-RAA fluorescence methods, TT values could be shortened by 1.72–2.32 and 2.60 minutes, respectively, and fluorescence values could be enhanced by 8847.25–9094.37 and 5250 mv, respectively. In the basic method, sensitivity could be increased by 10-fold. We successfully amplified a long-fragment of 509 bp in a P-RAA assay with a sensitivity of 102 copies/μL (compared with 103 copies/μL in the RAA assay). We thus conclude that betaine and pullulan are effective additives to enhance RAA assays.

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

2019 ◽  
Author(s):  
Andie C. Hall
Keyword(s):  
Nucleic Acid ◽  
Ethidium Bromide ◽  
Gel Electrophoresis ◽  
Careful Consideration ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Cost Increase ◽  
Dna Migration ◽  
Discernible Effect ◽  
Staining Methods

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%.

Luminography – an Alternative Assay for Detection of von Willebrand Factor Multimers

1988 ◽  
Vol 60 (02) ◽  
pp. 133-136 ◽  
Author(s):  
R Schneppenheim ◽  
H Plendl ◽  
U Budde
Keyword(s):  
Von Willebrand Factor ◽  
Gel Electrophoresis ◽  
Detection Methods ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Luminescence Assay ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryA luminescence assay was adapted for detection of von Willebrand factor multimers subsequent to SDS-agarose gel electrophoresis and electroblotting onto nitrocellulose. The method is as fast as chromogenic detection methods and appears to be as sensitive as autoradiography without the disadvantages of the latter.

Sign in / Sign up

Export Citation Format

Share Document