fuchsin (magenta I)
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This stain contains fuchsin or magenta I, a decolorized dye which has
a strong affinity for DNA, producing a red color in its presence. Also
called "rosaniline," is an aniline dye which consists of three aniline
rings, one meta-methylated, para-attached to a central carbon. It's positively
charged nitrogen is attracted to and stains the negatively charged PO4
groups in the DNA backbone.
It is nearly colorless until it reacts with the PO4 which turns it red. |
For 30 mL of the stain:
| STAGE | OPERATION |
| 1. Weigh | Weigh out 150 mg of Basic Fuchsin into 250 mL beaker. |
| 2. Dissolve | Pour 30 mL boiling dH2O over the Basic Fuchsin, swirl to dissolve. |
| 3. Cool and filter. | Cool to 50C, filter through Whatman No. 1 paper. |
| 4. Acidify | Add 4.5 mL 1N HCl, mix. (Caution: corrosive acid) |
| 5. Decolorize | Dissolve 0.45 g K2S2O5 (potassium
metabisulfite) to decolorize the solution.
Set in dark for 24 hours to bleach. |
| 6. Filter | .Decant or filter through activated charcoal (e.g., Norit) to further clarify (if desired). |
| 7. Storage | Store in light-proof bottle in refrigerator. (I.e. wrap in aluminum foil to exclude light.) |
| NOTE: |
It looks innocent because of its light straw color before it reacts with PO4 , but it will stain you seriously if you get in on your hands or clothes! |
[Fuchsin(e) Acid is related, but meta-sulfonated on all three aniline rings.]