We first obtained a nitrate broth tube and used the aseptic technique to inoculate the nitrate broth tube with a loopful of bacteria from our agar slant culture.
After two days, we added 5 drops of reagent A (sulfanilic acid) and 5 drops of reagent B (dimethyl-alpha-naphthylamine) to our inoculated tube. We made sure to wear gloves when we added the nitrate reagents.
After one to two minutes, we did not see a color change to pink or red color which means that we now need to add a small amount of powdered zinc to the tube.
After about 10 minutes we saw a small color change to our broth. This means that our bacteria are not able to reduce nitrate ions to either nitrite ions or to nitrogen gas.
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