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Zones of chitinase activity were observed after staining and destaining with 0.
Chandra TJ, Selvaraj R, Sharma YV Same day sputum smear microscopy with an economical destaining step for the diagnosis of pulmonary tuberculosis.
Slides were incubated for 1 h at room temperature in phosphoprotein stain (Invitrogen #MP33300; Thermo Fisher Scientific) while protected from light, destained for 3 x 30 min with a destaining solution specific to the stain (Invitrogen, #P33310; Thermo Fisher Scientific), and washed again for 2 x 10 min in ultrapure water.
The destaining time of Amaranth was much less than the routine dyes and quick results were obtained; given in Table-1.
Followed by staining with ethidium bromide (1 ug/mL) for 30 minutes and destaining with distilled water for 10 minutes.
After appropriate destaining with mixture of distilled water, methanol and acetic acid (9:6:1 v/v), the gels were photographed on Transilluminator.
The protein gels were fixed using destaining solution immediately after electrophoresis and the protein bands were visualized under UV light (290 nm).
Methylene blue is an alternative DNA stain that is noncarcinogenic but results in inferior staining and often requires more time for staining and destaining (Herrin and Schmidt, 1988).
Samples including both the low and high range molecular markers were diluted at different ratios 1:3, 1:4, 1:5 and 1:6 (v/v) in loading buffer and run at a constant voltage of 150 for an hour after which the gel was stained in coomasie brilliant blue and the molecular weights of the protein subunits determined after destaining for 2 hours, by a calibration graph obtained using the molecular weight markers.
5 mL of stain stock with 184 mL destaining solution (1:3:6 glacial acetic acid: methanol: distilled water) (12).
Gel electrophoresis procedures can be subject to variation due to differences in sample preparation, sample loading, staining and/or destaining time and image acquisition.
After the electrophoresis, the gel slab was incubated in dark with 50 ppm Sigma's Stains-All in 50% ethanol for 24 h, rinsed three times by water, incubated in dark with water for 24 h, and exposed to light for 2 h for further destaining.
The relative low cost of these dyes and their ready-made solutions, sensitivity in the 5-50 ng range and time-tested staining and destaining protocols have been fundamental to their wide acceptance.
After staining, the gel was washed twice in distilled water to remove excess of the stain then the gel was covered with 200-250 ml of destaining solution (45:45:10% Methanol: Acetic acid: Water).
The stain is inexpensive, is at least as sensitive as methylene blue, does not require destaining, requires visualisation with only visible light, and is specific for DNA but not RNA.
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