Negative staining with zinc-imidazole of gel electrophoresis-separated nucleic acids

Electrophoresis
E HardyL Castellanos-Serra

Abstract

Zinc and imidazole salts were applied for the detection of nucleic acids on either polyacrylamide of agarose gels. After electrophoresis, polyacrylamide gels are washed in distilled water to remove most of the residual electrophoresis reagents, then incubated in 10 mM zinc sulfate for 10 min, and subsequently immersed in 0.2 M imidazole for 3 min. As a result, zinc salts precipitate on the gel surface, except in the positions occupied by nucleic acids, which appear as transparent, colorless bands. Staining of nucleic acids on agarose gels can be performed by incubation in 40 mM zinc sulfate for 10 min, followed by immersion in 0.2 M imidazole for 5 min to form a deep white-stained background. On soaking in 2 M imidazole for 45 min, the imidazole-induced zinc precipitate is removed from the positions were nucleic acids are located resulting in a negative image of colorless and transparent nucleic acid bands against a white background. The sensitivity of this stain ranges from 5 to 7 ng/band for small (from 1 to 0.2 kbp) DNA, from 7.8 to 13 ng/band for different 22-base oligonucleotides, from 62 to 125 ng/band for large (from 20 to 2 kbp) DNA, and is 1 microgram/band for human peripheral-blood monocyte RNA. After chelation of zin...Continue Reading

Citations

Jun 21, 2011·Journal of Chemical Biology·Victoria J GauciJens R Coorssen
Apr 5, 2007·Nature Protocols·Lila Castellanos-Serra, Eugenio Hardy
Aug 10, 2000·Nucleic Acids Research·O Ihle, T E Michaelsen
Feb 8, 2000·Analytical Biochemistry·T M BrickerL K Frankel
Mar 9, 2010·Analytical Biochemistry·Karen LeónLilia López-Cánovas
Feb 20, 2007·Forensic Science International : Synergy·Lev A ZhivotovskyIosif S Tsybovsky
Apr 15, 2004·Analytical Biochemistry·Eugenio Hardy, Lila R Castellanos-Serra
May 30, 1998·Analytical Biochemistry·E HardyL R Castellanos-Serra
Mar 26, 1998·Analytical Biochemistry·L R Castellanos-SerraJ C Sánchez
Aug 31, 2021·Systems Biology in Reproductive Medicine·Kalpana JorasiaMeenaxi Sareen

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