Zinc-imidazole negative staining of chromosomal-sized DNA molecules in agarose minigels.
We standardized the zinc-imidazole negative staining method for detecting chromosomal-sized DNA molecules separated by pulsed field minigel electrophoresis. The best experimental conditions were as follows: separating large DNA molecules in minigels of 0.4 cm thickness, further incubating them with 40 mM ZnSO4 solution, and finally incubating them with 0.1 and 2 M imidazole solutions successively. The lowest yeast cells/miniplug useful in DNA band detection was 3 x 10(7) cells, as occurred with ethidium bromide-stained minigel. Electrophoresis patterns were visualized as colorless bands contrasting against a white background after illuminating the minigel with white light. This negative staining method is nontoxic and preserves the chemical integrity of the DNA molecules.
León
Karen
K
Molecular Biology Department, Cuban Neuroscience Center, Cubanacan, Playa, Ciudad Havana, Cuba.
Riverón
Ana María
AM
Arencibia
Oscar
O
López-Cánovas
Lilia
L
D004271:DNA, Fungal / Q000032:analysis*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D007093:Imidazoles; D016624:Negative Staining / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D015032:Zinc
D004271:DNA, Fungal; D007093:Imidazoles; C029899:imidazole; D015032:Zinc