Detection of DNA in Agarose Gels Using Berberine and Mordant Yellow 3R

Jung, Dawoon; Yoo, Gyurng-Soo; Choi, Jung-Kap

doi:10.1006/abio.1999.4198

Cited by 7 publications

(8 citation statements)

References 9 publications

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“…However, EB has several disadvantages in DNA staining because of having a strong mutagenic effect and causing damage to sample DNA and laboratory operators by UV illumination [33][34][35]. To avoid the strong mutagenic effect of EB, a fluorescent method using berberine/Mordant Yellow 3R (BB/M3R) has been published by our group [18]. The method has a comparable sensitivity of EB method, although it still requires UV irradiation.…”

Section: Staining Of Dna On Agarose Gels With Counterion Dyesmentioning

confidence: 97%

“…The mechanism of the counterion dye stain is also applicable to DNA staining on PAGE or agarose gels with other combinations of counterion dyes [18,20]. The principal staining mechanism of DNA is the same as that of protein but the roles of dye are the opposite to those of protein.…”

Section: Mechanism Of Counterion Dye Stain For Dnamentioning

confidence: 98%

“…This review focuses on the development of some visible dye stains studied in our laboratory, particularly the counterion dye staining methods for the analysis of protein [11][12][13][14][15][16][17] or DNA [18][19][20]. In addition, we also discuss a visible dye stain of protein on blotting membranes [21].…”

Section: Introductionmentioning

confidence: 99%

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Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Choi

2004

Electrophoresis

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Since 1993, we have studied visible organic dye stains for protein or DNA to improve methodologies and developed the counterion dye staining method. The method employs two oppositely charged dyes that form an ion-pair complex in the staining solution. The selective binding of free dye to protein or DNA in the staining solution improves detection sensitivity and speed. It is a rapid and sensitive procedure, involving fixing/staining or staining/quick destaining steps that are completed in 1-1.5 h. The lowest detection limits achieved are 4-8 ng of protein on polyacrylamide gels and approximately 10 ng of DNA on agarose gels. The focus of this review is to chronicle the development and current status of the counterion dye staining method for detection of protein or DNA. As an extended application of visible dyes, we also discuss the visible dye staining method for detecting protein on blotting membranes developed in our laboratory.

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Section: Staining Of Dna On Agarose Gels With Counterion Dyesmentioning

confidence: 97%

Section: Mechanism Of Counterion Dye Stain For Dnamentioning

confidence: 98%

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Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Choi

2004

Electrophoresis

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“…Additional studies have shown berberine to interact with diverse macromolecules such as liver alcohol dehydrogenase [73], human serum albumin [74], acetylcholinesterase [75], lipids [76], estrogens [77], Electrostatic interaction bile salts [78] and DNA [79]. The interaction of berberine with DNA [80][81][82][83][84] has been elucidated in fine detail including the effect of other molecules such as different metal cations and cyclodextrins [85,86].…”

Section: Biochemical Applicationsmentioning

confidence: 99%

Berberine: A Fluorescent Alkaloid with a Variety of Applications from Medicine to Chemistry

Domingo

Pardo²,

Cebolla

et al. 2010

MROC

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Berberine, a fluorescent, aromatic, heterocyclic alkaloid, produced in a diversity of plants, was found to have curative properties in Chinese herbal medicine. Berberine is used as a treatment for gastric and inflammatory diseases and as an antimicrobial agent. More recently the potential of this alkaloid in diverse new medical application like cardiology, neurology and oncology is being investigated. As a result of its fluorescent properties, it has also been used in chemistry to study protein conformation and protein/protein interactions and to quantitate non-fluorescent compounds. All these applications will be summarized and discussed in this review.

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“…However, EB has several disadvantages in staining DNA because of having a strong mutagenic effect and causing damages to sample DNA and laboratory personals by UV illumination [2][3][4]. To avoid the strong mutagenic effect of EB, a fluorescent method using berberine/mordant yellow 3R (BB/M3R) has been published by our group [5]. The method has a comparable sensitivity of EB method, although it still requires UV irradiation.…”

Section: Introductionmentioning

confidence: 99%

Counterion-dye staining method for DNA in agarose gels using crystal violet and methyl orange

Yang¹,

Jung²,

Bai

et al. 2001

Electrophoresis

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Sensitive and safe methods for visualization of DNA in agarose gels are described. 0.001% crystal violet dissolved in distilled water was used for DNA staining on agarose gels and it could detect as little as 16 ng of DNA (3 kb, pGem-7Zf/EcoRI) without destaining procedure. The detection limit is four times lower than that of ethidium bromide. To improve the sensitivity, we studied a counterion-dye staining method using methyl orange as a counterion-dye which contributes to reduce excessive background staining by crystal violet. Dye concentration, pH of staining solution, mixing molar ratio of two dyes, and staining times were optimized for the counterion-dye staining. By the staining with a mixed solution of 0.0025% crystal violet and 0.0005% methyl orange in distilled water, 8 ng of the 3 kb DNA in an agarose gel was detected within 30 min.

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Detection of DNA in Agarose Gels Using Berberine and Mordant Yellow 3R

Cited by 7 publications

References 9 publications

Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Berberine: A Fluorescent Alkaloid with a Variety of Applications from Medicine to Chemistry

Counterion-dye staining method for DNA in agarose gels using crystal violet and methyl orange

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