AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL

Objective: Present study was aimed to develop a reproducible, cheap and sensitive method for silver staining of double stranded DNA in agarose gel. Study Design: Experimental, repeated measure design. Place and Duration of Study: Department of Genetics, University of Karachi, Karachi. This...

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Autores principales:	Nuzhat Aisha Ikram, Bushra Iftikhar, Shoaib Ahmed, Fasiha Fatima, Tehmina Qamar, Shakeel R Farooqi
Formato:	article
Lenguaje:	EN
Publicado:	Army Medical College Rawalpindi 2019
Materias:	silver staining protocol dsdna Medicine R Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/405c7cfa7fb8410f9f1c7f89036e193d
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spelling	oai:doaj.org-article:405c7cfa7fb8410f9f1c7f89036e193d2021-11-16T03:00:01ZAN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL0030-96482411-8842https://doaj.org/article/405c7cfa7fb8410f9f1c7f89036e193d2019-02-01T00:00:00Zhttps://www.pafmj.org/index.php/PAFMJ/article/view/2502/2057https://doaj.org/toc/0030-9648https://doaj.org/toc/2411-8842Objective: Present study was aimed to develop a reproducible, cheap and sensitive method for silver staining of double stranded DNA in agarose gel. Study Design: Experimental, repeated measure design. Place and Duration of Study: Department of Genetics, University of Karachi, Karachi. This experimental study was conducted, from Nov 2013 to Jan 2014. Material and Methods: The new method is the modification and improvement in the original method proposed in the literature. Samples of ds genomic DNA was run on a nondenaturing 1.5% agarose/0.5X TBE. After electrophoresis gel was fixed in 10% acetic acid and staining was performed using 1 gm % silver nitrate. DNA bands were developed using 1.5% NaOH. At each step shaking was done manually with a circular movement. The modified method was also compared with the ethidium bromide staining of the same samples of DNA. Results: The modified method was proved to be as sensitive as the ethidium bromide with the advantage of having long term conservation ability of the gel. The main advantage of the protocol is the consumption of far less concentrations of silver nitrate and sodium hydroxide and the avoidance of the use of sodium thiosulphate. Conclusion: This method was easily reproducible, sensitive, and cheap with improved conservation ability of gel. It also avoids use of hazardous, expensive and time consuming radioactive and fluorescent detection.Nuzhat Aisha IkramBushra IftikharShoaib AhmedFasiha FatimaTehmina QamarShakeel R FarooqiArmy Medical College Rawalpindiarticlesilver staining protocoldsdnaMedicineRMedicine (General)R5-920ENPakistan Armed Forces Medical Journal, Vol 69, Iss 1, Pp 87-91 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	silver staining protocol dsdna Medicine R Medicine (General) R5-920
spellingShingle	silver staining protocol dsdna Medicine R Medicine (General) R5-920 Nuzhat Aisha Ikram Bushra Iftikhar Shoaib Ahmed Fasiha Fatima Tehmina Qamar Shakeel R Farooqi AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL
description	Objective: Present study was aimed to develop a reproducible, cheap and sensitive method for silver staining of double stranded DNA in agarose gel. Study Design: Experimental, repeated measure design. Place and Duration of Study: Department of Genetics, University of Karachi, Karachi. This experimental study was conducted, from Nov 2013 to Jan 2014. Material and Methods: The new method is the modification and improvement in the original method proposed in the literature. Samples of ds genomic DNA was run on a nondenaturing 1.5% agarose/0.5X TBE. After electrophoresis gel was fixed in 10% acetic acid and staining was performed using 1 gm % silver nitrate. DNA bands were developed using 1.5% NaOH. At each step shaking was done manually with a circular movement. The modified method was also compared with the ethidium bromide staining of the same samples of DNA. Results: The modified method was proved to be as sensitive as the ethidium bromide with the advantage of having long term conservation ability of the gel. The main advantage of the protocol is the consumption of far less concentrations of silver nitrate and sodium hydroxide and the avoidance of the use of sodium thiosulphate. Conclusion: This method was easily reproducible, sensitive, and cheap with improved conservation ability of gel. It also avoids use of hazardous, expensive and time consuming radioactive and fluorescent detection.
format	article
author	Nuzhat Aisha Ikram Bushra Iftikhar Shoaib Ahmed Fasiha Fatima Tehmina Qamar Shakeel R Farooqi
author_facet	Nuzhat Aisha Ikram Bushra Iftikhar Shoaib Ahmed Fasiha Fatima Tehmina Qamar Shakeel R Farooqi
author_sort	Nuzhat Aisha Ikram
title	AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL
title_short	AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL
title_full	AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL
title_fullStr	AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL
title_full_unstemmed	AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL
title_sort	improved modified protocol for silver staining of ds dna in agarose gel
publisher	Army Medical College Rawalpindi
publishDate	2019
url	https://doaj.org/article/405c7cfa7fb8410f9f1c7f89036e193d
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AN IMPROVED MODIFIED PROTOCOL FOR SILVER STAINING OF DS DNA IN AGAROSE GEL

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