Gram Staining

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Recommended: Microbiology lab exam gram stain

The samples unknow bacteria from cut toenail swabbed and inoculate a tube of Tryptone Soy Broth (TSB). The tube at 30o C for 24 to 36 hours with shaked. When it passed 24 to 36 hours then perform Gram stain to find identify Gram positive and negative. Obtain a BBLTM Gram slide with ten squares with two different squares marked C for control of Gram positive and negative. Smear a small amount of bacteria from toenail in TSB into squares #1 and waited for the slide to completely dry. Then added a drop of methanol to each square on the slide and to get rid of methanol and allowed it get dry. The slide got dried then added flood on the slide with crystal violet stain and allowed for 30 seconds and both Gram positive and negative which it was Gram …show more content…

Inoculate a tube unknow of TSB to tryptophan TSB. All of tubes and plates placed at 30oC for two days.
After two days and observed tubes and plates and record the result. Thioglycolate tube and observed of bacteria growth throughout the tube at the top, bottom, or both.
Fermentation broths and observed for turbidity in the tube showed positive or negative for growth. When it changed color which into acid and if the durham tube with a bubble, it showed that the bacterium produced H2 production.
The streak on the TSA plate and observed with flood the plate with iodine and allowed the plate for about 2 minutes. If it showed a clear zone around streak then it was a positive result.
The isolated colonies on TSA and record the result if they all look generally the same or not. Then do catalase test to showed a drop of catalase reagent on a colonies to see gas bubbles appeared which it showed a positive test. The motility medium to observe the growth only along the stab line or grew out from the stab line the resulted in an uniformly turbid tube.
The indole test by added 10 drops of BBLTM Indole reagent to TSB. If TSB showed a red ring at the top of the broth and it was positive test for

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